We thank all residents and attending physicians, notably Dr. A.F. Haney (the University of Chicago, Department of Obstetrics and Gynecology) for collecting omental biopsies. Also, we thank Stacey Tobin and Gail Isenberg for carefully editing this manuscript. This work was supported by Bears Care, the charitable beneficiary of the Chicago Bears Football Club, the National Institute of Neurological Disorders and Stroke (NINDS) R21 NS075702, and the National Cancer Institute grant R01 CA111882 to E.L.

Tarif edilen tüm araştırma protokolleri Chicago Kurumsal Değerlendirme Kurulu Üniversitesi&#39;nde (KİK) tarafından incelenmiştir. Aydınlatılmış onam ameliyattan önce her hastadan alındı ​​ve çalışma Chicago IRB Üniversitesi tarafından onaylanmıştır. Korunması için insan dokusu taşıma ve hücrelerin kontamine riskini azaltmak için ne zaman bir biyolojik güvenlik kabini tip 2 ve eldiven kullanılmalıdır. Birincil dönüştürülmemiş Stromal Hücreleri 1. İzolasyon ve Kültür <ol><li> İnsan doku toplama ve hazırlama. <ol><li> Abdominal operasyon esnasında alınan insan omentum, 2 cm3, örneklerini elde edilir ve hemen RT fosfat tamponlu tuzlu su (PBS) (Şekil 2A), dokuyu bırakın. X 2 cm omentum 3 cm&#39;lik bir parçası, tipik olarak 1.000.000 birincil insan mezoteliyal hücreleri (HPMC) ve 200,000 primer insan fibroblastları ve normal omental fibroblastlar (NOF) verir. </li><li> Doku 3 için 0.5 xg&#39;de dalmış PBS aşağı Spindak kısa sürede sonra toplanması (&lt;2 PBS içinde saat) ve 50 ml konik bir tüp içinde 20 ml taze PBS doku aktarın. 15 cm çapında, steril kültür çanak (Şekil 2B) neşter ile kıyma ile 5mm 3 parçaya doku yukarı kesti. </li></ol></li><li> Birincil insan mezotel hücre izolasyonu 8 <ol><li> HPMC izole etmek için, 50 ml konik tüpler içine kıyılmış doku transferi ve katı parçaları üst (Şekil 2C-D) float için 1 dakika bekleyin. HPMC ve kırmızı kan hücrelerinin (RBC) alt kısmında sıvı içinde mevcut olacaktır. Borunun alt ve yeni bir konik tüp içine sıvının çıkması için bir pipet kullanın. 3 dakika boyunca 0.5 xg sıvıyı aşağı Spin ve HPMC / RBC pelet süpernatant aspire. <ol><li> , Kıyılmış dokuya 20 ml PBS eklemek doku, aşağı spin, yükselmeye bir pipet ile ve HPMC / RBC tüpüne alttan sıvıyı çıkarmak için izin ve kaldırın: işlemini üç kez tekrarlayınsüpernatant. Tüm spin tamamlanır ve süpernatan aspire edildikten sonra, pelet HPMC ve RBC&#39;yi içerecektir. </li></ol></li><li> Plaka% 10 cenin sığır serumu [FBS],% 1 MEM vitamin [93 mg / L],% 1 MEM esansiyel olmayan amino asitler [81,4 tam büyüme ortamı, 15 ml DMEM (75 cm 2 şişe içine pelet tüm hücrelerin mg / L],% 1 penisilin streptomisin [Pen-strep, 100 U / ml penisilin ve 100 ug / ml streptomisin]). </li><li> Ek HPMCs izole etmek için ikinci bir PBS yıkama yapın. <ol><li> Doku üstüne yüzen ikincil PBS yıkama için, 200 rpm&#39;de, 20 ml PBS içinde 10 dakika süreyle 37 ° C &#39;de geri kalan katı doku sallayın, daha sonra 1 dakika bekleyin. 3 dakika boyunca 0,5 x g&#39;de santrifüj, yeni bir tüp içine tüpün altındaki sıvıyı çıkarın ve süpernatan aspire. </li><li> Tam büyüme ortamı 15 ml ayrı 75 cm 2 şişe yıkama, ikincil PBS tüm HPMC / RBC Plate. </li></ol></li><li> Herhangi r izole etmek içinhacimce PBS solüsyonu: 1,% 0.25 tripsin, 25 mM EDTA: HPMC emaining, 1, 20 ml, 10 dakika boyunca, 200 rpm&#39;de 37 ° C derece kıyılmış doku sallayın. Doku yükselmeye tüpün alt kısmında sıvı toplamak, yeni bir 50 ml tüp içine ve 0.5 gx 3 dakika aşağı dönmesine izin verin. <ol><li> Süpernatant aspire tam büyüme ortamında 15 ml ayrı 75 cm 2 şişe tüm HPMC / RBC plaka. </li></ol></li><li> Kültür nemlendirilmiş bir ortamda 37 ° C&#39;de ve% 5 CO2 plakalandı hücreleri. Harcanan medya çıkarmadan günlerde 3 ve 5 tam büyüme ortamı 15 ml ekleyerek Yem kaplı hücreler. HPMC bölme önce 5-7 gün süre ile kültüre edilebilir. <ol><li> Orijinal fenotip dediferansiyonunu ve modifikasyonu en aza indirmek için tüm deneylerde (geçit 2 kadar) düşük geçit HPMC&#39;ye kullanın. Faz kontrast capabilitie plastik diferansiyel girişim kontrast yetenekleri veya 4-20x amacı ile 20x objektif ile tercihen geniş alan mikroskobu kullanıns hücrelerinin tüm görüntüleri çekmek için (mikroskop faz kontrast filtreleri) ve 3,3&#39;-diaminobenzidin (DAB) boyama immunohistokimyasal analiz parlak alanında 10x objektif (). </li></ol></li><li> HPMC cuboidal olduğunu onaylayın ve immunohistokimyasal 8 (Şekil 3A, C, E) gerçekleştirerek sitokeratinin 8 ve vimentinin ifade eder. </li></ol></li><li> NOF izolasyonu 8 <ol><li> 1: 100 x Pen-Strep 1 karışımı bir 1: birleştirerek kollajenaz tip III çözeltisi (10 x) &#39;in bir stokunun 10 ml hazırlayın kollajenaz tip III 1500 ünite aktivite / ml PBS içinde hyaluronidaz 714 ünite aktivite / ml olmuştur. </li><li> % 1 MEM vitamin [93 mg / L] ile serumsuz ortam içinde seyreltilmiş 10 kat kollajenaz tip III çözeltisi, 10 ml (DMEM 6 saat süre ile 200 rpm&#39;de HPMC izolasyon sonra kalan kıyılmış omentum doku 37 ° C sallayın, % 1 MEM esansiyel olmayan amino asitler [81,4 mg / L],% 1 penisilin streptomisin [Pen-strep, 100 U / ml penisilin ve 100 ug / ml streptomisin]). Sindirilmiş doku opak bakacağız ve bazı lifli enkaz (Şekil 2E) olabilir. </li><li> Çözelti içeren NOF hücreleri Santrifüj oda sıcaklığında 3 dakika boyunca 0,5 x g &#39;de süspansiyon halinde, süpernatan aspire tam büyüme ortamında 13 ml bir 75 cm&#39;lik 2 şişeye pelet plaka. </li><li> Eski ortamı çıkarın ve 24 saat sonra taze tam büyüme ortamı 15 ml ile değiştirin. NOF bölme önce 1-3 gün süre ile kültive edilebilir. Hücreler konfluense ulaştığınızda NOF notu çoğalması sona erecek. </li><li> Birincil insan fibroblastlar, düz, uzun hücreler olduğunu onaylayın ve vimentinin ifade ama immünohistokimyasal 8 gerçekleştirerek 8 sitokeratin değil (Şekil 3B, D, F). Dediferansiyonunu en aza indirmek için (ideal geçit 3 öncesi) deneyler için düşük geçit NOF kullanın ve orijinal fenotip modifikasyonu. Hücrelerin her aşaması fotoğraf çekmek için plastik DIC yetenekleri ile 20x amacı ile geniş alan mikroskobu kullanınve parlak alanında 10x objektif immunohistokimyasal DAB boyama analiz etmek. </li></ol></li></ol> 2. Organotipik Kültür 8 Kaplama <ol><li> En fazla 5 dakika boyunca 3 ml% 0.25 tripsin / EDTA 25 mM, ardından 10 ml PBS ile çalkalayın 75 cm kültür şişesi Yayın NOF. Tam büyüme ortamının en az 3 kez hacmi ile nötralize tripsin. </li><li> 50 ml konik tüp içine tripsinize hücreleri aktarın ve 0.5 gx 3 dk aşağı doğru döndürün. Süpernatantı ve tam büyüme ortamı 5 ml kadar geri hücreleri getirmek. Kültür şişesi kurtarıldı hücreleri saymak için bir hücre sayacı kullanın. </li><li> Bir 96-yuvalı, berrak tabanlı siyah plaka üzerine 100 ul başına 2000-4000 NOF plaka tam büyüme ortamı, uygun hacimde hücreleri ile seyreltilir. Kaplama önce hücre karışıma sıçan kuyruğu kollajen I 0.5 ug / 100 ul ekleyin. Let Hücrelerin en az 4 saat boyunca nemlendirilmiş bir ortamda% 5 CO2 içinde 37 ° C &#39;de oturup, ya da hücrelerin kadarplaka yüzeyine yapışır için. </li><li> Adımlar 2.1 ve 2.2 açıklanan aynı yöntemleri kullanarak kültür şişesi HPMC bırakın. Tam büyüme ortamının uygun miktarda hücrelerin önceden 96 oyuklu plakalar içinde kollajen I kaplama NOF üstünde 50 ul başına 10,000-20,000 HPMC plaka seyreltin. Hücreler deney başlamadan önce en az 18 saat boyunca nemlendirilmiş bir ortamda% 5 CO2 içinde 37 ° C &#39;de bekletin. </li><li> Tam büyüme ortamı HeyA8 yumurtalık kanser hücreleri -expressing Kültür yeşil flüoresan proteini (GFP). HeyA8 yumurtalık kanseri hücreleri, floresan kopepod cGFP (pCDH-CMV-MCS-EF1) eksprese eden bir lentiviral vektör ekspresyonu ile etiketlenir. HeyA8 yumurtalık kanseri hücreleri, kullanım gününde (logaritmik büyüme fazı) üzerine% 80-90 ortak akışlı olmalıdır. Kültür hücreleri plakadan bırakın ve birincil hücreler için adımlar 2,1-2,2 açıklanan aynı yöntemleri kullanarak saymak. </li><li> Yumurtalık kanseri hücreleri, 37 ° C&#39;de 15-20 dakika boyunca tam bir büyüme ortamı içinde kurtarmak için izin verinkapaklı konik tüp gevşek kapatılmış. </li></ol> 3. Yapışma Deneyi 8 <ol><li> İyileştikten sonra 0,5 xg&#39;de 3 dakika boyunca iplik ile yumurtalık kanseri hücrelerini pellet haline getirmek ve daha sonra süpernatant kaldırmak ve 50.000 hücre / 100 ul bir konsantrasyona ulaşmak için serum içermeyen ortam, uygun hacimde hücreleri seyreltin. </li><li> Harcanan ortam kaldırmak için HPMC / NOF Organotipik kültürler ile 96 oyuklu plakalar ters çevirin, ve her bir serumsuz ortam içinde kanser hücre süspansiyonu 100 ul ilave edin. 4 saat 30 dakika boyunca nemlendirilmiş bir ortamda% 5 CO2 içinde 37 ° C &#39;de plaka inkübe edin. </li><li> Ortam ve yapışmayan hücreleri çıkarmak için 96 oyuklu plaka ters çevirin. Dikkatle ve yavaşça her kuyuya PBS 100 ul pipetlemeyin ve tekrar plaka ters düşük güçte bir çok kanallı pipet kullanın. PBS bir kez daha yıkayın tekrarlayın. </li><li> PBS çıkarın ve her bir oyuğa 100 ul% 4 paraformaldehid eklemek için ters çevirin. Plaka 20 mil için bekletinn hücreleri gidermek için. 20 dakika sonra, plaka ters çevirin ve 100 ul PBS ilave edin. </li><li> Toplam kuyu flüoresan kaydedilmemiştir veya hücre sayısı belirlenebilir unutmayın. Ölçümü için, 1.000.000 hücre / ml&#39;lik bir konsantrasyonda HeyA8 hücreleri kullanılarak, iki kopya halinde birinci plaka standart eğrisi. <ol><li> Aşağı 1 milyon hücre iplik medya kaldırarak, ve onları 20 dakika boyunca 1 ml% 4 paraformaldehid oturup izin vererek standart eğri olun. </li><li> Spin hücreleri tekrar PBS (anlattıklarında hücreleri 1000000 / ml konsantrasyon onaylamak için) ml 1&#39;de getirmek. Levha 50.000, 40.000, 30.000, 20.000, 10.000, 5.000, 1.000, ve de her biri çifte biçimde içine 0 hücreleri, ve her çukur 50 ul&#39;lik bir nihai toplam hacim PBS ilave edin. </li></ol></li><li> Alt yüksek kuyu (standart eğri üzerinde 50.000) ölçekleme, bir flüoresan plaka okuyucusu (uyarım = 488 nm, emisyon = 528 nm) ile kültür plakasına okuyun. Organotyp yapıştırılır kaç yumurtalık kanseri hücrelerini hesaplamak için standart eğri kullanınHer bir oyuğa (Şekil 4A-C) &#39;ic kültürü. </li></ol> 4. Proliferasyon Deneyi 10 <ol><li> Yumurtalık kanseri hücreleri, geri sonra% 1 FBS (DMEM,% 1 FBS ortamı uygun hacimde hücreleri sulandırmak sonra 0.5 x g&#39;de 3 dakika boyunca iplik hücreleri pelet,% 1 MEM (adım 2.5 ve yukarıda belirtilen 2,6 bakınız) vitaminler [93 mg / L],% 1 MEM esansiyel olmayan amino asitler [81,4 mg / L],% 1 penisilin streptomisin [Pen-Strep, 100 U / ml penisilin ve 100 ug / ml streptomisin]) 4.000 hücre konsantrasyonu elde etmek için / 150 ul. </li><li> Her bir oyuğa FBS ortamı harcanan ortamını çıkarın ve 1% kanser hücre süspansiyonu 150 ul eklemek için HPMC / NOF Organotipik kültürler ile 96 oyuklu plaka ters çevirin. 72 saat boyunca nemlendirilmiş bir ortamda% 5 CO2 içinde 37 ° C &#39;de plaka inkübe edin. </li><li> Alt görece proliferasyonunu değerlendirmek için bir kez bir flüoresan plaka okuyucusu günde (uyarım = 488 nm, emisyon = 528 nm) kültür plaka okumahücre. 48 saat (Şekil 5A-C) &#39;de ortamının değiştirilmesi, 96 saat boyunca deney devam edin. (Plaka tercih edilir tersini) ortam değiştirirken kültürünü rahatsız etmemek için dikkatli olun. </li></ol> 5. Invasion Testi 8 <ol><li> 3D kültür kaplama önce, 24 oyuklu bir kültür plakasına parçası (8 um gözenek boyutu) 200 ul PBS içinde 7.5 ug fare kuyruk kolajeni eklemek. Plaka inkübe O / N, daha sonra dikkatlice hemen kollajen kaplı ekler (yukarıdaki adım 2) üzerine HPMC / NOF organotipik kültürünü kaplama önce bir pipet ile PBS kaldırmak edelim. </li><li> Adım 3.1 olarak yumurtalık kanseri hücrelerini hazırlayın. Ekler üzerinde organotipik kültürlerinin üzerine yumurtalık kanseri hücrelerini plaka dikkatle ekler üst fazla olan ortamı çıkarmak için bir pipet kullanın. Her bir oyuğa, serumsuz ortam içinde yumurtalık kanseri hücre süspansiyonu 100 ul ekle. </li><li> Kanser hücresini ikna etmek için iyi bir ekin altındaki içine tam büyüme ortamı 750 ul ekleyinorganotipik kültürlerinin içine vasion. 24 saat boyunca nemlendirilmiş bir ortamda% 5 CO2 içinde 37 ° C &#39;de plaka inkübe edin. Not: Organotipik kültür ekleme çapraz ve ekin alt tarafında kalacaktır istila Yumurtalık kanseri hücrelerini. </li><li> Boş kuyuya insert taşıyın, 24 saat sonra, maşa ile ekleme kavramak ve kısaca PBS beher durulayın. 1 dakikalık bir toplam pamuklu çubuk her iki yanına, her bir ek üst fýrçalayýn. Hızlı bir şekilde her bir 750 ul% 4 paraformaldehid içeren bir plaka içine ekleme yerleştirin. Her insert 750 ul PBS ile dolu kuyu içine ekler aktarmadan önce 10 dakika boyunca paraformaldehid oturup bekleyin. </li><li> 2.5x büyütme mikroskop ile işgal hücreler (ekler dibine yapıştırılır ve sabit) görüntüleyin. Tüm kuyu görüş alanı içinde olduğu zaman, kaçınarak, 10x büyütme ayarlayın ve 5 resim, kuyunun her kadranda merkezine yakın bir ve 1 almakkenarlarına çok yakın görüntüler alarak. Her iyi için aynı desen izleyin. </li><li> Onların protokole göre her bir kuyu (Şekil 6A-C) &#39;de alan başına işgal hücrelerin ortalama sayısını elde etmek için her görüntüdeki hücre sayısını saymak için üreticinin programı kullanın. </li></ol>

<ol>
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Metastasis. 5, 181-197 (1987).</li><li>Kanemoto, T., Martin, G. R., Hamilton, T. C., Fridman, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+synthetic+peptides+and+protease+inhibitors+on+the+interaction+of+a+human+ovarian+cancer+cell+line+(NIH:OVCAR-3)+with+a+reconstituted+basement+membrane+(matrigel).">Effects of synthetic peptides and protease inhibitors on the interaction of a human ovarian cancer cell line (NIH:OVCAR-3) with a reconstituted basement membrane (matrigel).</a> Invasion Metastasis. 11, 84-92 (1991).</li><li>Rieppi, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesothelial+cells+induce+the+motility+of+human+ovarian+carcinoma+cells.">Mesothelial cells induce the motility of human ovarian carcinoma cells.</a> Int. J. Cancer. 80, 303-307 (1999).</li><li>Barbolina, M. V., Adley, B. P., Ariztia, E. V., Liu, Y., Stack, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microenvironmental+regulation+of+membrane+type+1+matrix+metalloproteinase+activity+in+ovarian+carcinoma+cells+via+collagen-induced+EGR1+expression.">Microenvironmental regulation of membrane type 1 matrix metalloproteinase activity in ovarian carcinoma cells via collagen-induced EGR1 expression.</a> J. Biol. Chem. 282, 4924-4931 (2007).</li><li>Burleson, K. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ovarian+carcinoma+ascites+spheroids+adhere+to+extracellular+matrix+components+and+mesothelial+cell+monolayers.">Ovarian carcinoma ascites spheroids adhere to extracellular matrix components and mesothelial cell monolayers.</a> Gynecol. Oncol. 93, 170-181 (2004).</li><li>Suzuki, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HMOCC-1,+a+human+monoclonal+antibody+that+inhibits+adhesion+of+ovarian+cancer+cells+to+human+mesothelial+cells.">HMOCC-1, a human monoclonal antibody that inhibits adhesion of ovarian cancer cells to human mesothelial cells.</a> Gynecol. Oncol. 95, 290-298 (2004).</li><li>Kishikawa, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+distinct+pattern+of+peritoneal+involvement+shown+by+in+vitro+and+in+vivo+ovarian+cancer+dissemination+models.">Two distinct pattern of peritoneal involvement shown by in vitro and in vivo ovarian cancer dissemination models.</a> Invas. Metast. 15, 11-21 (1995).</li><li>Casey, R. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+an+in+vitro+assay+to+measure+the+invasion+of+ovarian+carcinoma+cells+through+mesothelial+cell+monolayers.">Establishment of an in vitro assay to measure the invasion of ovarian carcinoma cells through mesothelial cell monolayers.</a> Clin. Exp. Metastasis. 20, 343-356 (2003).</li><li>Niedbala, M. J., Crickard, K., Bernacki, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interactions+of+human+ovarian+tumor+cells+with+human+mesothelial+cells+grown+on+extracellular+matrix.">Interactions of human ovarian tumor cells with human mesothelial cells grown on extracellular matrix.</a> Exp. Cell Res. 160, 499-513 (1985).</li><li>White, E. A., Kenny, H. A., Lengyel, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-Dimensional+modeling+of+ovarian+cancer.">Three-Dimensional modeling of ovarian cancer.</a> Adv. Drug Deliv. Rev. 79-80, 184-192 (2014).</li><li>Kenny, H. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+high+throughput+screening+using+a+primary+human+three-dimensional+organotypic+culture+predicts+in+vivo+efficacy.">Quantitative high throughput screening using a primary human three-dimensional organotypic culture predicts in vivo efficacy.</a> Nature Comm. , (2015).</li><li>Sawada, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=c-Met+overexpression+is+a+prognostic+factor+in+ovarian+cancer+and+an+effective+target+for+inhibition+of+peritoneal+dissemination+and+invasion.">c-Met overexpression is a prognostic factor in ovarian cancer and an effective target for inhibition of peritoneal dissemination and invasion.</a> Cancer Res. 67, 1670-1680 (2007).</li><li>Kenny, H. A., Lengyel, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MMP-2+functions+as+an+early+response+protein+in+ovarian+cancer+metastasis.">MMP-2 functions as an early response protein in ovarian cancer metastasis.</a> Cell cycle. 8, 683-688 (2009).</li><li>Sawada, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Loss+of+E-cadherin+promotes+ovarian+cancer+metastasis+via+alpha+5-integrin,+which+is+a+therapeutic+target.">Loss of E-cadherin promotes ovarian cancer metastasis via alpha 5-integrin, which is a therapeutic target.</a> Cancer Res. 68, 2329-2339 (2008).</li><li>Mitra, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ligand-independent+activation+of+c-Met+by+fibronectin+and+&#945;(5)&#946;(1)-integrin+regulates+ovarian+cancer+invasion+and+metastasis.">Ligand-independent activation of c-Met by fibronectin and &#945;(5)&#946;(1)-integrin regulates ovarian cancer invasion and metastasis.</a> Oncogene. 30, 1566-1576 (2011).</li></ol>

The authors have nothing to disclose.

Periton mikroçevresinin bir Organotipik modeli yumurtalık kanseri yayılmasına mikroçevresinin hücresel ve doğuştan bileşenlerin hem bireysel ve kolektif işlev (ler) değerlendirmek için kurulmuştur. Yumurtalık kanseri hücre yapışması, proliferasyon ve istila araştırmak için 3D Organotipik kültürü kaplama ve uyarlama için özel protokolleri temin edilmektedir. Primer insan omental mezotel hücreleri ve fibroblastlar hastalardan izole edilen ve normal morfoloji ve hasta varyasyonu korumak için erk...

Ovarian cancer is the deadliest gynecologic malignancy1. The majority of patients are diagnosed after the cancer has disseminated throughout the peritoneal cavity. Once the cancer has spread throughout the peritoneal cavity, cytoreductive surgery and chemotherapy are often not sufficient treatment to prevent cancer recurrence and chemoresistance, resulting in a less than 30% 5-year survival rate. Ovarian cancer metastasis is predominantly limited to the peritoneal cavity, and several other cancer types, including gastric, pancreatic, and colon cancers, metastasize to the same anatomic sites in the peritoneal cavity. In general, ovarian cancer cells detach from the in situ carcinoma in the fallopian tube or the primary ovarian tumor, travel in peritoneal fluid as single cells or spheroids, and attach to mesothelium-lined surfaces of the omentum, bowel, and abdominal wall2. 
The tumor microenvironment plays an important role in disease progression and chemoresistance in many cancers3-6. The peritoneal cavity is a unique microenvironment, with a mesothelial cell monolayer covering the majority of surfaces (Figure 1A)7. The mesothelial lining acts as a barrier that creates a low-friction surface, which tends to be protective against cancer cell adhesion8. Immediately underneath this mesothelial-lined surface is a layer made predominantly of fibroblasts and extracellular matrix (ECM), which promote cancer cell adhesion and invasion8. Ovarian cancer cells secrete factors that induce changes in the mesothelial cell lining that enhance ovarian cancer cell adhesion, invasion, and metastasis9,10. Ovarian cancer cells adhere to the mesothelial surface via integrin and CD44-mediated mechanisms (Figure 1B)11-16. 
Historically, several 3D models have been developed to investigate ovarian cancer interactions with the microenvironment. Some of the first models studied ovarian cancer-ECM interface17-21, ovarian cancer-mesothelial cell communication13,14,21-24, or both25 (reviewed by us 26). Niedbala et al. discovered that ovarian cancer cells display a quicker and firmer adhesion to ECM than to mesothelial cells or to plastic alone25. However, these models did not histologically resemble the peritoneal microenvironment. Therefore, we established a 3D organotypic model to more thoroughly replicate the ovarian cancer microenvironment. In order to better understand the role of the microenvironment and the interaction between cancer and peritoneal cells in the peritoneal dissemination of ovarian cancer, we have developed a 3D organotypic in vitro culture model of the peritoneal cavity lining (schematic in Figure 1C). The proposed model is composed of primary human fibroblasts and ECM, covered with a layer of primary human mesothelial cells-each cell type is isolated from human omentum. Histologically, this model resembles the normal peritoneal or omental lining, and provides a surface on which we can study the tumor microenvironment, the interaction between cancer cells and normal tissue, and the processes of cancer cell adhesion, invasion, and proliferation8.

<table><tbody><tr><td>&lt;strong&gt;1. Isolation and culture of primary cells&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>PBS</td><td>Fisher Scientific</td><td>SH3001304</td><td></td></tr><tr><td>Single-edged razor blades</td><td>Fisher Scientific</td><td>12-640</td><td></td></tr><tr><td>15 cm culture dishes</td><td>BD Biosciences</td><td>353025</td><td></td></tr><tr><td>Glass flask</td><td>?</td><td>?</td><td></td></tr><tr><td>Fetal Bovine Serum (FBS)</td><td>Life Technologies</td><td>16000044_3616914956</td><td></td></tr><tr><td>DMEM with L-Glutamine</td><td>Corning</td><td>10-013-CV</td><td></td></tr><tr><td>MEM Vitamins</td><td>Corning</td><td>25-020-Cl</td><td></td></tr><tr><td>MEM Nonessential amino acids</td><td>Corning</td><td>25-025-CI</td><td></td></tr><tr><td>Penicillin-Streptomycin</td><td>Corning</td><td>30-002-CI</td><td></td></tr><tr><td>Shaker&amp;nbsp;</td><td>Thermo-Fisher</td><td>MaxQ 4450</td><td></td></tr><tr><td>Centrifuge</td><td>Eppendorf</td><td>5702</td><td></td></tr><tr><td>Incubator</td><td>Thermo-Fisher</td><td>Forma Series II Water Jacketed CO2 Incubator Model 3100</td><td></td></tr><tr><td>Trypsin EDTA, 1x (0.25%)</td><td>Corning</td><td>25-053-CI</td><td></td></tr><tr><td>Hyaluronidase</td><td>Worthington Biochemical</td><td>LS002592</td><td></td></tr><tr><td>T-75 Flasks</td><td>BD Biosciences</td><td>353136</td><td></td></tr><tr><td>T-175 Flasks</td><td>BD Biosciences</td><td>353112</td><td></td></tr><tr><td>Pipet tips</td><td>Rainin</td><td>P2, P10, P20, P200 and P1000</td><td></td></tr><tr><td>Pipet tips</td><td>Corning</td><td>Filtered tips P2, P10, P20, P200 and P1000</td><td></td></tr><tr><td>&lt;strong&gt;Name of Reagent/ Equipment&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments/Description&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;2. Plating 3D culture&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Cell Counter</td><td>Invitrogen</td><td>Countess</td><td></td></tr><tr><td>Countess Cell Counting Chamber Slides</td><td>Invitrogen</td><td>C10313</td><td></td></tr><tr><td>Trypan Blue Stain (0.4%)</td><td>Gibco</td><td>15250-061</td><td></td></tr><tr><td>Collagen Type I (Rat Tail)</td><td>BD Biosciences</td><td>354236</td><td></td></tr><tr><td>96 well plate, clear bottom, black</td><td>BD Biosciences</td><td>353219</td><td></td></tr><tr><td>&lt;strong&gt;Name of Reagent/ Equipment&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments/Description&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;3. Adhesion assay&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Multichannel pipet</td><td>Eppendorf</td><td>Xplorer 300</td><td></td></tr><tr><td>Paraformaldehyde solution 4% in PBS</td><td>Santa Cruz Biotechnology</td><td>sc-281692</td><td></td></tr><tr><td>Plate reader</td><td>Molecular Devices</td><td>Minimax</td><td></td></tr><tr><td>&lt;strong&gt;Name of Reagent/ Equipment&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments/Description&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;5. Invasion assay&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Cell Culture Inserts (8um, 24-well)</td><td>BD Biosciences</td><td>353097</td><td></td></tr><tr><td>Cotton swabs</td><td>Q-tips</td><td>cotton swabs</td><td></td></tr><tr><td>Microscope</td><td>Zeiss</td><td>Axiovert 200m</td><td></td></tr><tr><td>Cell Profiler</td><td>public domain</td><td></td><td></td></tr><tr><td>24 well plate</td><td>BD Biosciences</td><td>353047</td><td></td></tr><tr><td>&lt;strong&gt;Name of Reagent/ Equipment&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments/Description&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;6. Antibodies&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Anti-Integrin &amp;alpha;V&amp;beta;3 Antibody, clone LM609</td><td>EMD Millipore</td><td>MAB1976</td><td></td></tr><tr><td>Beta 1&amp;nbsp;</td><td>Oncosynergy</td><td>OS2966</td><td></td></tr><tr><td>Alpha 5 [CD49e]</td><td>ID Pharmingen</td><td>555615</td><td></td></tr><tr><td>Beta 4 [CD104]</td><td>EMD Millipore</td><td>MAB 2058</td><td></td></tr></tbody></table>

modeling the early steps of ovarian cancer dissemination in an organotypic culture of the human peritoneal cavity

The pattern of ovarian cancer metastasis is markedly different from that of most other epithelial tumors, because it rarely spreads hematogenously. Instead, ovarian cancer cells exfoliated from the primary tumor are carried by peritoneal fluid to metastatic sites within the peritoneal cavity. These sites, most notably the abdominal peritoneum and omentum, are organs covered by a mesothelium-lined surface. To investigate the processes of ovarian cancer dissemination, we assembled a complex three-dimensional culture system that reconstructs the lining of the peritoneal cavity in vitro. Primary human fibroblasts and mesothelial cells were isolated from human omentum. The fibroblasts were then mixed with extracellular matrix and covered with a layer of the primary human mesothelial cells to mimic the peritoneal and omental surfaces encountered by metastasizing ovarian cancer cells. The resulting organotypic model is, as shown, used to examine the early steps of ovarian cancer dissemination, including cancer cell adhesion, invasion, and proliferation. This model has been used in a number of studies to investigate the role of the microenvironment (cellular and acellular) in early ovarian cancer dissemination. It has also been successfully adapted to high throughput screening and used to identify and test inhibitors of ovarian cancer metastasis.

Organotipik kültürü ilk olarak kollajen tip I primer insan fibroblastları karıştırılması ve sonra mezotelyal hücrelerin 5 kez sayı ile bu kültürü overlaying tarafından toplandı. Yumurtalık kanseri hücreleri, yapışma, istilası veya proliferasyonunu çalışma ilave edilmeden önce, kültür en az 18 saat süreyle inkübe edildi. Her bir deney, çoklu tekrarlanmıştır (n = 5) ve istila deneyler (n = 3) (n = 3-5), farklı hastalara ve çok sayıda kuyu elde edilen 3D kültürleri yapışma her durumd...

Watch this Scientific Journal Video about Modeling the Early Steps of Ovarian Cancer Dissemination in an Organotypic Culture of the Human Peritoneal Cavity at JoVE.com

Modeling the Early Steps of Ovarian Cancer Dissemination in an Organotypic Culture of the Human Peritoneal Cavity

Here, we present a protocol to construct a three-dimensional in vitro model of the lining of the peritoneal cavity, composed of primary human mesothelial cells and fibroblasts layered with extracellular matrix, as a tool to investigate ovarian cancer cell adhesion, invasion, and proliferation.

İnsan periton boşluğuna bir Organotipik Kültür Yumurtalık Kanseri Yaygınlaştırılması Erken Adımlar Modelleme

The pattern of ovarian cancer metastasis is markedly different from that of most other epithelial tumors, because it rarely spreads hematogenously. ...

modeling-early-steps-ovarian-cancer-dissemination-an-organotypic

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Medicine

26.5K Görüntüleme.  University of Chicago. Here, we present a protocol to construct a three-dimensional in vitro model of the lining of the peritoneal cavity, composed of primary human mesothelial cells and fibroblasts layered with extracellular matrix, as a tool to investigate ovarian cancer cell adhesion, invasion, and proliferation. 

Video: Modeling the Early Steps of Ovarian Cancer Dissemination in an Organotypic Culture of the Human Peritoneal Cavity

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

Epithelial ovarian carcinoma (EOC) is associated with a poor prognosis because it shows peritoneal dissemination. To improve the prognosis, it is important to control peritoneal dissemination. However, it is still unclear how tumor cells detach from primary lesions and attach to the mesothelium. The establishment of an appropriate animal model is needed to gain an understanding of the mechanism of peritoneal dissemination in vivo. In the current study, we introduce the process from the local injection of EOC cells into the murine ovarian surface to the development of metastasis, including the peritoneum and distant organs. Female nude mice (BALB/c nu/nu) at 8 weeks of age were used. Under a microscopic field of view, EOC cells (1 x 105 cells/&#181;l of medium-extracellular matrix (ECM)-based hydrogel/unilateral ovary/mouse) were injected into murine ovaries through a retroperitoneal approach from the dorsal flank. This proposed method is a less invasive procedure for the mouse and minimizes damage to the ovary. Here, we describe the methodological steps in the development of the original and metastatic tumor formation of EOC.

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

To clarify the multistep process of peritoneal dissemination of ovarian carcinoma, the current study presents a murine experimental model of original tumor development and peritoneal metastasis via orthotopic inoculation with these tumor cells.

Orijinal Tümör Geliştirme Murin Deneysel Modeli ve Periton Metastaz<em&gt; yoluyla</emYumurtalık Kanseri Hücreleri ile&gt; Ortotopik aşılama

Epithelial ovarian carcinoma (EOC) is associated with a poor prognosis because it shows peritoneal dissemination. To improve the prognosis, it is ...

Kanser Araştırmaları

In vitro Mesothelial Clearance Assay that Models the Early Steps of Ovarian Cancer Metastasis

Ovarian cancer is the fifth leading cause of cancer related deaths in the United States1. Despite a positive initial response to therapies, 70 to 90 percent of women with ovarian cancer develop new metastases, and the recurrence is often fatal2. It is, therefore, necessary to understand how secondary metastases arise in order to develop better treatments for intermediate and late stage ovarian cancer. Ovarian cancer metastasis occurs when malignant cells detach from the primary tumor site and disseminate throughout the peritoneal cavity. The disseminated cells can form multicellular clusters, or spheroids, that will either remain unattached, or implant onto organs within the peritoneal cavity3 (Figure 1, Movie 1). 
All of the organs within the peritoneal cavity are lined with a single, continuous, layer of mesothelial cells4-6 (Figure 2). However, mesothelial cells are absent from underneath peritoneal tumor masses, as revealed by electron micrograph studies of excised human tumor tissue sections3,5-7 (Figure 2). This suggests that mesothelial cells are excluded from underneath the tumor mass by an unknown process. 
Previous in vitro experiments demonstrated that primary ovarian cancer cells attach more efficiently to extracellular matrix than to mesothelial cells8, and more recent studies showed that primary peritoneal mesothelial cells actually provide a barrier to ovarian cancer cell adhesion and invasion (as compared to adhesion and invasion on substrates that were not covered with mesothelial cells)9,10. This would suggest that mesothelial cells act as a barrier against ovarian cancer metastasis. The cellular and molecular mechanisms by which ovarian cancer cells breach this barrier, and exclude the mesothelium have, until recently, remained unknown. 
Here we describe the methodology for an in vitro assay that models the interaction between ovarian cancer cell spheroids and mesothelial cells in vivo (Figure 3, Movie 2). Our protocol was adapted from previously described methods for analyzing ovarian tumor cell interactions with mesothelial monolayers8-16, and was first described in a report showing that ovarian tumor cells utilize an integrin &#x2013;dependent activation of myosin and traction force to promote the exclusion of the mesothelial cells from under a tumor spheroid17. This model takes advantage of time-lapse fluorescence microscopy to monitor the two cell populations in real time, providing spatial and temporal information on the interaction. The ovarian cancer cells express red fluorescent protein (RFP) while the mesothelial cells express green fluorescent protein (GFP). RFP-expressing ovarian cancer cell spheroids attach to the GFP-expressing mesothelial monolayer. The spheroids spread, invade, and force the mesothelial cells aside creating a hole in the monolayer. This hole is visualized as the negative space (black) in the GFP image. The area of the hole can then be measured to quantitatively analyze differences in clearance activity between control and experimental populations of ovarian cancer and/ or mesothelial cells. This assay requires only a small number of ovarian cancer cells (100 cells per spheroid X 20-30 spheroids per condition), so it is feasible to perform this assay using precious primary tumor cell samples. Furthermore, this assay can be easily adapted for high throughput screening.

In vitro Mesothelial Clearance Assay that Models the Early Steps of Ovarian Cancer Metastasis

The mesothelial clearance assay described here takes advantage of fluorescently labeled cells and time-lapse video microscopy to visualize and quantitatively measure the interactions of ovarian cancer multicellular spheroids and mesothelial cell monolayers. This assay models the early steps of ovarian cancer metastasis.

In vitro Mezotelyal Gümrükleme Assay o Yumurtalık Kanseri Metastaz Erken Adımlar modeller

Ovarian cancer is the fifth leading cause of cancer related deaths in the United States1. Despite a positive initial response to therapies, 70 to 90 ...

Tıp

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical treatments and improved platinum-based chemotherapies, there has been little improvement in EOC survival rates for more than four decades 1,2. Whilst stage I tumors have 5-year survival rates &gt;85%, survival rates for stage III/IV disease are &lt;40%. Thus, the high rates of mortality for EOC could be significantly decreased if tumors were detected at earlier, more treatable, stages 3-5. At present, the molecular genetic and biological basis of early stage disease development is poorly understood. More specifically, little is known about the role of the microenvironment during tumor initiation; but known risk factors for EOCs (e.g. age and parity) suggest that the microenvironment plays a key role in the early genesis of EOCs. We therefore developed three-dimensional heterotypic models of both the normal ovary and of early stage ovarian cancers. For the normal ovary, we co-cultured normal ovarian surface epithelial (IOSE) and normal stromal fibroblast (INOF)&nbsp;cells, immortalized by retrovrial transduction of the catalytic subunit of human telomerase holoenzyme (hTERT) to extend the lifespan of these cells in culture. To model the earliest stages of ovarian epithelial cell transformation, overexpression of the CMYC oncogene in IOSE cells, again co-cultured with INOF cells. These heterotypic models were used to investigate the effects of aging and senescence on the transformation and invasion of epithelial cells. Here we describe the methodological steps in development of these three-dimensional model; these methodologies aren't specific to the development of normal ovary and ovarian cancer tissues, and could be used to study other tissue types where stromal and epithelial cell interactions are a fundamental aspect of the tissue maintenance and disease development.

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

We describe methodologies for establishing in vitro heterotypic three-dimensional models comprising ovarian fibroblasts and normal ovarian surface or ovarian cancer epithelial cells. We discuss the use of these models to study stromal-epithelial interactions that occur during ovarian cancer development.

Üç boyutlu heterotipik<em&gt; In Vitro</emYumurtalık Kanseri Başlatma ve İlerleme sırasında Stromal-Epitel Etkileşimleri&gt; Modelleme

Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical ...

Assessment of Ovarian Cancer Spheroid Attachment and Invasion of Mesothelial Cells in Real Time

Ovarian cancers metastasize by shedding into the peritoneal fluid and dispersing to distal sites within the peritoneum. Monolayer cultures do not accurately model the behaviors of cancer cells within a nonadherent environment, as cancer cells inherently aggregate into multicellular structures which contribute to the metastatic process by attaching to and invading the peritoneal lining to form secondary tumors. To model this important stage of ovarian cancer metastasis, multicellular aggregates, or spheroids, can be generated from established ovarian cancer cell lines maintained under nonadherent conditions. To mimic the peritoneal microenvironment encountered by tumor cells in vivo, a spheroid-mesothelial co-culture model was established in which preformed spheroids are plated on top of a human mesothelial cell monolayer, formed over an extracellular matrix barrier. Methods were then developed using a real-time cell analyzer to conduct quantitative real time measurements of the invasive capacity of different ovarian cancer cell lines grown as spheroids. This approach allows for the continuous measurement of invasion over long periods of time, which has several advantages over traditional endpoint assays and more laborious real time microscopy image analyses. In short, this method enables a rapid, determination of factors which regulate the interactions between ovarian cancer spheroid cells invading through mesothelial and matrix barriers over time.

Ovarian cancer cell invasion into the mesothelial lining of the peritoneum is a dynamic process over time. Utilizing a real time analyzer, the invasive capacity of ovarian cancer cells in a spheroid-mesothelial cell co-culture model can be quantified over prolonged time periods, providing insights into factors regulating the metastatic process.

Real Time Mezotelyal Hücrelerinin Yumurtalık Kanseri Sferoid Ek ve Invasion Değerlendirilmesi

Ovarian cancers metastasize by shedding into the peritoneal fluid and dispersing to distal sites within the peritoneum. Monolayer cultures do not ...

Over Kanseri-Omentum Etkileşimi için 3 Boyutlu Ex Vivo Model Oluşturulması

An Ex Vivo Model of Ovarian Cancer Peritoneal Metastasis Using Human Omentum

Ovarian cancer is the deadliest gynecologic malignancy. The omentum plays a key role in providing a supportive microenvironment to metastatic ovarian cancer cells as well as immune modulatory signals that allow tumor tolerance. However, we have limited models that closely mimic the interaction between ovarian cancer cells and adipose-rich tissues. To further understand the cellular and molecular mechanisms by which the omentum provides a pro-tumoral microenvironment, we developed a unique 3D ex vivo model of cancer cell-omentum interaction. Using human omentum, we are able to grow ovarian cancer cells within this adipose-rich microenvironment and monitor the factors responsible for tumor growth and immune regulation. In addition to providing a platform for the study of this adipose-rich tumor microenvironment, the model provides an excellent platform for the development and evaluation of novel therapeutic approaches to target metastatic cancer cells in this niche. The proposed model is easy to generate, inexpensive, and applicable to translational investigations.

Yazar Spotlight: Kanser-Adipoz Mikroçevre Etkileşimini Araştırmak için Gelişmiş Ex Vivo Model 

This protocol describes the establishment of a three-dimensional (3D) ex vivo model of cancer cell-omentum interaction. The model provides a platform for elucidating pro-tumor mechanisms within the adipose niche and for testing novel therapies.

İnsan Omentumu Kullanılarak Over Kanseri Periton Metastazının Ex Vivo Modeli

Ovarian cancer is the deadliest gynecologic malignancy. The omentum plays a key role in providing a supportive microenvironment to metastatic ovarian ...

Modeling Ovarian Cancer Multicellular Spheroid Behavior in a Dynamic 3D Peritoneal Microdevice

Ovarian cancer is characterized by extensive peritoneal metastasis, with tumor spheres commonly found in the malignant ascites. This is associated with poor clinical outcomes and currently lacks effective treatment. Both the three-dimensional (3D) environment and the dynamic mechanical forces are very important factors in this metastatic cascade. However, traditional cell cultures fail to recapitulate this natural tumor microenvironment. Thus, in vivo-like models that can emulate the intraperitoneal environment are of obvious importance. In this study, a new microfluidic platform of the peritoneum was set up to mimic the situation of ovarian cancer spheroids in the peritoneal cavity during metastasis. Ovarian cancer spheroids generated under a non-adherent condition were cultured in microfluidic channels coated with peritoneal mesothelial cells subjected to physiologically relevant shear stress. In summary, this dynamic 3D ovarian cancer-mesothelium microfluidic platform can provide new knowledge on basic cancer biology and serve as a platform for potential drug screening and development.

To study ovarian tumor progression in a physiologically relevant model, multicellular spheroids were cultured in a microdevice under simulated fluid flow. This dynamic 3D model emulates the intraperitoneal environment with the cellular and mechanical components where ovarian cancer metastasis occurs.

Dinamik 3D Periton microdevice Modelleme Yumurtalık Kanseri Multisellüler Sfero Davranış

Ovarian cancer is characterized by extensive peritoneal metastasis, with tumor spheres commonly found in the malignant ascites. This is associated with ...

Biyomühendislik

Modeling Ovarian Cancer Communication for Imaging Mass Spectrometry Analysis: A Method to Identify Small Molecule Chemical Communicators in Metastatic Ovarian Tumor Development

Source: Zink, K. E. et. al. <a href="https://www.jove.com/t/59490">Capturing Small Molecule Communication Between Tissues and Cells Using Imaging Mass Spectrometry.</a> J. Vis. Exp. (2019)
This protocol describes a technique for sample preparation by co-culturing ovarian explants with the fallopian tube cancer cells. These samples are visualized using imaging mass spectroscopy, or IMS,&#160; to detect small molecule exchange between the cells and tissues leading to tumor development.

Görüntüleme Kütle Spektrometresi Analizi için Over Kanseri İletişiminin Modellenmesi: Metastatik Over Tümörü Gelişiminde Küçük Moleküllü Kimyasal İletişimcileri Tanımlamak İçin Bir Yöntem

Source: Zink, K. E. et. al. Capturing Small Molecule Communication Between Tissues and Cells Using Imaging Mass Spectrometry. J. Vis. Exp. (2019)
This ...

JoVE Encyclopedia of Experiments

Jinekolojik Kanser

Analizler ve teknikler

Orthotopic Inoculation of Ovarian Cancer Cells into Murine Ovary: A Procedure to Establish an Ovarian Cancer Mouse Model to Study Peritoneal Dissemination of Cells

Source: Koya, Y. et al. <a href="https://www.jove.com/t/54353">Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells.</a> J. Vis. Exp. (2016)
This video describes the protocol for creating a murine experimental model to study tumor development and peritoneal metastasis. This ovarian cancer model is generated by the orthotopic inoculation of human epithelial ovarian cancer cells into the ovaries of a female mouse.

Yumurtalık Kanseri Hücrelerinin Murin Yumurtalığına Ortotopik Aşılanması: Hücrelerin Peritoneal Yayılımını İncelemek için Bir Yumurtalık Kanseri Fare Modeli Oluşturma Prosedürü

İnsan periton boşluğuna bir Organotipik Kültür Yumurtalık Kanseri Yaygınlaştırılması Erken Adımlar Modelleme

Özet

Daha Fazla Video Keşfet

Bu videodaki bölümler

Orijinal Tümör Geliştirme Murin Deneysel Modeli ve Periton Metastaz

In vitro Mezotelyal Gümrükleme Assay o Yumurtalık Kanseri Metastaz Erken Adımlar modeller

Üç boyutlu heterotipik

Real Time Mezotelyal Hücrelerinin Yumurtalık Kanseri Sferoid Ek ve Invasion Değerlendirilmesi

İnsan Omentumu Kullanılarak Over Kanseri Periton Metastazının Ex Vivo Modeli

İnsan Omentumu Kullanılarak Over Kanseri Periton Metastazının Ex Vivo Modeli

İnsan Omentumu Kullanılarak Over Kanseri Periton Metastazının Ex Vivo Modeli

Dinamik 3D Periton microdevice Modelleme Yumurtalık Kanseri Multisellüler Sfero Davranış

Görüntüleme Kütle Spektrometresi Analizi için Over Kanseri İletişiminin Modellenmesi: Metastatik Over Tümörü Gelişiminde Küçük Moleküllü Kimyasal İletişimcileri Tanımlamak İçin Bir Yöntem

Yumurtalık Kanseri Hücrelerinin Murin Yumurtalığına Ortotopik Aşılanması: Hücrelerin Peritoneal Yayılımını İncelemek için Bir Yumurtalık Kanseri Fare Modeli Oluşturma Prosedürü