Destek: NIDCD R01DC009801, P30DC010362, 5T32DC000040-17

Etik Açıklama: Bu protokolde insan tümör örneklerinin kullanılması Iowa Kurumsal Değerlendirme Kurulu Üniversitesi (KİK) tarafından onaylanmıştır. 1.. Kur Gün önce Tümör Hasat <ol><li> Coat Hücre Kültürü Plakaları: bir steril kültür başlık olarak, kapak yerine, bir polistiren / plastik kültür kuyu / çanak alt karşılamak için yeterli poli-L-ornitin (% 0.01 solüsyon) ekleyin ve en azından oda sıcaklığında bekletin 2 hr. Cam slaytlar, lameller veya görüntüleme gerektiren deneyler için polistiren için ikame edilebilir. </li><li> 2 saat sonra, emme ile poli-L-ornitin çözeltisi çıkarın ve plakaları bir steril kültür davlumbaz tamamen kurumaya bırakın. Kadar laminin solüsyonu (Ca 2 10 ug / ml + / Mg2 + içermeyen HBSS [HBSS-/ -]) tamamen kültür plakaları kaplamak için, kapak yerine, daha sonra gece boyunca 4 ° C soğuk buzdolabı içinde 1) yeri, ya ya da 2), en az 2 saat boyunca 37 ° C inkübatör yerleştirin. Eğer plakalargecede veya daha uzun saklanır, buharlaşma / kuruma önlemek için plastik wrap plakaları sarın. </li></ol> 2.. Kurulum Tümör Hasat Günü <ol><li> Hazırlama Kültür Ortamı: Ortam, tümör hasat ve işleme gününde taze yapılmış 4 ° C&#39;de saklanır ve hemen önce ortam ilave ve / değişir inkübatörde 37 ° C&#39;ye kadar ısıtılır. </li><li> Schwannom kültür ortamı 0,1 mg / ml penisilin ve 0.1 mg / ml streptomisin ile yüksek glikoz (4.5 mg / ml) (fenol kırmızısı ile) DMEM olduğu; N2 ortamı son seyreltme 1:100 takviyesi; Bovine İnsülin 5 ug / ml; % 10 toplam hacme Fetal Sığır Serumu. 0.22 mikron filtre ile tüm medya filtre. </li><li> Örnek Transport Soğutucu hazırlayın: ameliyathaneye 2 + (HBSS + / +) 2 + Ca / Mg ile birlikte 25 ml HBSS ile steril 50 ml konik tüp (hasat edilmesi, tümörün miktarına bağlı olarak) 1-2, bir buz dolu bir soğutucu gönder tümör numune transferi ve ulaşım için. </li><li> Indiv hazırlayınidual tümör ayrışma tüpleri - 1 HBSS&#39;nin + / + ml ve buz üzerinde yer ile yuvarlak steril 2 ml alt tüpler doldurun. (Tüpler tümör örneklerinin keskin diseksiyon / ayrışması için kullanılan edilecektir). </li><li> Ayrışma için, hasat edilen doku, her 0.5 cm 3 için yaklaşık bir ayrılma tüpü hazırlanması; örneğin, 1.0 x 1.0 x 1.0 cm (1 cm3) ölçen bir tümör numunesi iki HBSS + / + ayrılma tüpler gerektirecektir. </li><li> Ultraviyole ışık sterilizasyon: Yer doku makas, küçük forseps, neşter sapı, 1x normal bir Petri kabı, P1000 pipet ve steril bir kültürü kaputu pipet uçları, ve ~ için ultraviyole ışık altında bırakmak öncesinde tümör dokusunun geliş ve işleme 15 dk. </li></ol> 3.. Numune İzolasyon ve Ulaşım <ol><li> Cerrahi rezeksiyon ile tümör örnekleri izole. </li><li> Hemen hastadan çıkarıldıktan sonra mümkün olan en kısa sürede, buzla soğutulmuş HBSS + / + içine tümör örnekleri yerleştirin. Bir kez tüm tissue örnekleri daha fazla işlem için laboratuara ameliyat tiyatro, (buz üzerinde) taşıma örnekleri toplanır. </li></ol> 4.. Doku Ayrılma ve triturasyon <ol><li> Laminer akış kaputu standart, steril tekniği izleyerek, kültürü kaputu içine tümör örneği ile konik tüp getirmek ve tüp ve tümör 50x evirerek örnekleri yıkayın. Fragmanları, tüpün altına düşer ve daha sonra HBSS + / + çıkarmak için izin verin. 30 ml HBSS + / + ile doldurun ve ajitasyon / tekrar 50x ters. 4x toplam inversiyon / medya kaldırma tekrarlayın. </li><li> 30 ml HBSS + / + son kez, sonra bir sterile100 mm petri içine karışımı dökün, ve ayrı tümör 0,5 cm içine 3 gruplar içinde tümör fragmanları askıya yeniden. Büyük tümör fragmanları karşılaşılan, küçük parçalar halinde kesmek için makas veya neşter doku (# 11 veya # 15 bıçak) kullanın. </li><li> Bireysel HBSS + / + dolu içine 0.5 cm 3 doku grupları yerleştirmek için forseps kullanın, 2 ml&#39;lik yuvarlak dipli konikTüm buz üzerinde arkadaşları borular. A &#39;snowglobe&#39; görünümü (Şekil 1) sahip olmalıdır alt 1 mm fragmanları-süspansiyona tümör parçaları kıyma doku makas kullanın. Parçalarının çoğunluğu alt 1 mm kadar sadece tümör kıyma; değil &#39;over-kıyma&#39; tümör örnekleri yapın. Geri buz tamamen kıyılmış doku tüpü yerleştirin ve tüm ek numune tüpleri ile tekrarlayın. </li><li> Tek bir 15 ml konik tüp içine tüm parçalanmış tümör örnekleri aktarın. Bir kesim / modifiye steril ucu ile bir P1000 pipet, bu adım için iyi çalışır. Kullanın ekstra HBSS + / + emin tüm tümör numune 15 ml tüp içinde tahsil edilmiştir yapmak için 2 ml ayrışma tüpler durulama / temizlemek için. 5 dakika boyunca 73 x g, 23 ° C &#39;de santrifüj içinde bir karışımı aşağıya doğru dönerler. </li><li> Hücre topaklarını bırakarak HBSS + / + kapalı emme. % 0.25 tripsin 1:1 karışımı içinde fragmanları yeniden askıya (HBSS-/ içinde çözülmüş -) -% 0.2 &#39;lik kolajenaz tümör parçalarını tutmak için yeterince sıkı eklemek SUspension. 30 dakika boyunca 37 ° C kuluçka makinesi içinde vidalı üst ve bir yer değiştirme. Süspansiyon parçaları korumak için inkübasyon sırasında en az bir kez hücre karışımı çalkalayın. De bu anda ısınmak için kuvöz içine Schwannom kültür ortamı yerleştirin. </li><li> İnkübasyondan sonra, 23 ° C, 5 dakika için 73 x g tripsin etkisiz hale getirmek için her bir tüpe 100 ul FBS ekleme ve daha sonra santrifüj örnek. Bir pipet kullanarak, dikkatli bir hücre pelet bozmadan mümkün olduğunca süpernatant aspire. 01:02 tümör parçalarının nihai oranı tümör fragmanları için ısıtıldı kültür ortamı (N2/insulin/5% FBS) ekleyin kültür ortamı (örneğin tümör parçalarının 1 mi var ise, ısıtılmış ve 2 ml ekle medya). </li><li> Yaklaşık 1-2 mm&#39;lik bir çapa bir P1000 pipet ucu kapalı ucu kesilerek tümör toz haline getirme için hazırlayın. Kolayca bir unalt yoluyla çözüm pipetle kadar yavaş yavaş, giderek daha küçük ve daha küçük çaplı pipet ipuçlarını kullanarak hücre süspansiyonu çiğnemekniyor P1000 pipet. Kolayca ucundan tümör çözüm pipetle bir kez hemen küçük bir ucuna hareket - over örnek çiğnemek yok. Konik tüpün alt pipet çekme toz haline getirme sırasında doku blokları aspirasyon daha büyük tek bir parça, genellikle sürekli ince toz haline koyma izin veriyorsa. </li></ol> 5.. Hücre Kaplama <ol><li> Doku biri 0.5 mm 3 parçası 1 x 100 mm tabak veya 4 x 8-well/4-well slaytlar tohum için yeterlidir. Hemen önce medya / hücre karışımı ekleyerek, kültür plakaları laminin çözüm kaldırmak ancak plakaları kuru izin vermeyin. </li><li> Her 100 mm tabak için 10 mi ısıtıldı kültür ortamı olarak tümör parçası çözülür. </li><li> 4-iyi bir slayt her bir kuyu için 750 ul ısıttı kültür ortamı (tüm slayt için 3 ml) kullanın. </li><li> 8-iyi slayt her bir kuyu için 400 ul ısıttı kültür ortamı (tüm slayt için 3.2 ml) kullanın. </li><li> 37 ve en kültürleri inkübe# 160; ° C,% 100 nem,% 6 CO2. En az 36 saat süre ile, tek başına kültürleri bırakın ve daha sonra solüsyon (asidik) sararmış ise ortamı değiştirmek. Aksi takdirde daha sonra her 2-3 gün veya ortam asidik olunca, 72. saatte ortamı değiştirmek. Ortam üst üste birkaç gün boyunca günlük olarak asidik hale gelmesi durumunda, bu genellikle potansiyel bakteri ya da maya kontaminasyon bir göstergesidir. </li></ol>

<ol>
	<li>Arthurs, B. J., 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=A+review+of+treatment+modalities+for+vestibular+schwannoma.">A review of treatment modalities for vestibular schwannoma.</a> Neurosurg Rev. 34 (3), 265-277 (2011).</li><li>Evans, G. R., Lloyd, S. K., Ramsden, R. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurofibromatosis+type+2.">Neurofibromatosis type 2.</a> Adv Otorhinolaryngol. 70, 91-98 (2001).</li><li>Fong, B., 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=The+molecular+biology+and+novel+treatments+of+vestibular+schwannomas.">The molecular biology and novel treatments of vestibular schwannomas.</a> J Neurosurg. 115 (5), 906-914 (2011).</li><li>Jacob, 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=Preclinical+validation+of+AR42,+a+novel+histone+deacetylase+inhibitor,+as+treatment+for+vestibular+schwannomas.">Preclinical validation of AR42, a novel histone deacetylase inhibitor, as treatment for vestibular schwannomas.</a> Laryngoscope. 122 (1), 174-189 (2012).</li><li>Evans, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neurofibromatosis+2+[Bilateral+acoustic+neurofibromatosis,+central+neurofibromatosis.">Neurofibromatosis 2 [Bilateral acoustic neurofibromatosis, central neurofibromatosis.</a> NF2, neurofibromatosis type II]. Genet Med. 11 (9), 599-610 (2009).</li><li>Anniko, M., Noren, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Human+Acoustic+Neurinoma+in+Organ-Culture+.1.+Methodological+Aspects.">The Human Acoustic Neurinoma in Organ-Culture .1. Methodological Aspects.</a> Acta Oto-Laryngologica. 91 (1-2), 47-53 (1981).</li><li>Manent, J., 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=Magnetic+cell+sorting+for+enriching+Schwann+cells+from+adult+mouse+peripheral+nerves.">Magnetic cell sorting for enriching Schwann cells from adult mouse peripheral nerves.</a> J Neurosci Methods. 123 (2), 167-173 (2003).</li><li>Nair, S., 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=Primary+cultures+of+human+vestibular+schwannoma:+selective+growth+of+schwannoma+cells.">Primary cultures of human vestibular schwannoma: selective growth of schwannoma cells.</a> Otol Neurotol. 28 (2), 258-263 (2007).</li><li>Baur, A. 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=Laminin+promotes+differentiation,+adhesion+and+proliferation+of+cell+cultures+derived+from+human+acoustic+nerve+schwannoma.">Laminin promotes differentiation, adhesion and proliferation of cell cultures derived from human acoustic nerve schwannoma.</a> Acta Otolaryngol. 115 (4), 517-521 (1995).</li><li>Cravioto, H., 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=Experimental+neurinoma+in+tissue+culture.">Experimental neurinoma in tissue culture.</a> Acta Neuropathol. 21 (2), 154-164 (1972).</li><li>Kaasinen, S., 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=Culturing+of+acoustic+neuroma--methodological+aspects.">Culturing of acoustic neuroma--methodological aspects.</a> Acta Otolaryngol Suppl. 520 Pt 1, 25-26 (1995).</li><li>Kredel, F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue+Culture+of+Intracranial+Tumors+with+a+Note+on+the+Meningiomas.">Tissue Culture of Intracranial Tumors with a Note on the Meningiomas.</a> Am J Pathol. 4 (4), 337-340 (1928).</li><li>Murray, M. R., Stout, A. P., Bradley, C. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Schwann+cell+versus+fibroblast+as+the+origin+of+the+specific+nerve+sheath+tumor:+Observations+upon+normal+nerve+sheaths+and+neurilemomas+in+vitro.">Schwann cell versus fibroblast as the origin of the specific nerve sheath tumor: Observations upon normal nerve sheaths and neurilemomas in vitro.</a> Am J Pathol. 16 (1), 41-60 (1940).</li><li>Cravioto, H., Lockwood, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+behavior+of+acoustic+neuroma+in+tissue+culture.">The behavior of acoustic neuroma in tissue culture.</a> Acta Neuropathol. 12 (2), 141-157 (1969).</li><li>Lee, J. D., 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=Genetic+and+epigenetic+alterations+of+the+NF2+gene+in+sporadic+vestibular+schwannomas.">Genetic and epigenetic alterations of the NF2 gene in sporadic vestibular schwannomas.</a> PLoS One. 7 (1), 30418 (2012).</li><li>Wippold, F. J., 2nd, , 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=Neuropathology+for+the+neuroradiologist:+Antoni+A+and+Antoni+B+tissue+patterns.">Neuropathology for the neuroradiologist: Antoni A and Antoni B tissue patterns.</a> AJNR Am J Neuroradiol. 28 (9), 1633-1638 (2007).</li><li>Mennel, H. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short-term+tissue+culture+observations+of+experimental+primary+and+transplanted+nervous+system+tumors.">Short-term tissue culture observations of experimental primary and transplanted nervous system tumors.</a> J Neuropathol Exp Neurol. 39 (6), 639-660 (1980).</li><li>Rosenbaum, 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=Isolation+and+characterization+of+Schwann+cells+from+neurofibromatosis+type+2+patients.">Isolation and characterization of Schwann cells from neurofibromatosis type 2 patients.</a> Neurobiol Dis. 5 (1), 55-64 (1998).</li></ol>

Çıkar çatışması ifşa etmek.

In vitro Schwannom Kültürleri Tarihi Vitro schwanom kültürlerde kurmak için çabalar açık cerrahi rezeksiyon 1894 12 yılında meydana gelen ilk akustik nöroma sonra sadece bir kaç yıl başladı. İlk kaydedilen kültürleri başarısız &quot;asılı damla yöntemiyle&quot; 12 ile kıyılmış tümör büyümeye çalışmış 1920&#39;lerde Kredel tarafından vardı . Daha sonra, Dr. Murray ve Stout pıhtılaşmış...

Vestibüler schwannomalar (VSS) tüm intrakraniyal tümörlerin 1-3 10% ödün, Schwann hücresi vestibüler sinirin (SC) tümörler temsil eder. Vss hem TNF2 tümör baskılayıcı olarak inaktive kusurları ile ilişkili sporadik veya ailesel ya (Nörofibromatozis tip 2, NF2) formlar, meydana Gen. VSS tedavisi ancak bu tür prosedürlerin morbidite sağırlık, yüz nöropatlien omurilik sıvısı kaçağı, dengesizlik, ve tümör büyütme 1 içerir, genellikle cerrahi rezeksiyon veya radyocerrahi olduğunu. Buna ek olarak her hasta olarak kabul edilebilir bir cerrahi veya radyasyon adaylardır. Alternatif tedaviler gibi önemli morbidite yanı sıra bir eksikliği yeni tedaviler 3,4, gelişmekte umuduyla VSS eşsiz moleküler biyoloji soruşturma tahrik vardır. Hücre kültürü potansiyel tedavi com moleküler ve hücresel davranış ve tarama, hızlı kolay ve derinlemesine analiz sağlarlira. Tarihsel olarak, taze doku örneklerinin erişim ve schwannoma hücreler immortalize değildir gerçeğinin olmaması önemli schwannoma Tümörgenez araştırılması için primer kültürlerinde kullanımını engellemiştir. Primer kültürlerin sınırlı kaynağı üstesinden gelmek için, ölümsüz kılınmış hücre kuşağı HEI193 VS HPV E6 ve E7 onkogenler 5 ile transdüksiyonu ile üretilmiştir. Bu onkojenik transdüksiyon schwannoma, insan tümörleri için bir model olarak, kullanımlarını sınırlayan hücrelere, önemli moleküler ve fenotipik değişiklikler getirmiştir. Fonksiyonel NF2 geni eksik transgenik fare hatları türetilen SC in vitro kültürler NF2 bağımlı SC tümörogenez araştırmak için başka bir alternatif oluşturmaktadır. Bu kültürler, ancak insan belirli bir davranış insan VSS veya hesabın heterojen doğasını özetlemek için başarısız. Çoğu önceki VS kültür teknikleri nispeten uzun işlem süreleri ve karmaşık seçici kültür teknikleri 6-8 gereklidir.Immun boyama ile saptandığı haliyle Burada% 95 tümör hücresi saflıkta altında, 3 saat içinde tam bir işlem ile hücre kültürü VS primer için basit ve tekrarlanabilir bir protokol, mevcut.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Poly-L-Ornithine, 0.01% Solution</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">P4957</td>
			<td style="width:167px;">Cell Culture Surface Treatment</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Laminin Mouse Protein&#160;</td>
			<td style="width:71px;">Gibco</td>
			<td style="width:119px;">23017-015 / L2020</td>
			<td>Cell Culture Surface Treatment</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">0.2% Collagenase (dissolved in HBSS -/-)</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">C2674</td>
			<td>Dissociation Reagent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">0.25% Trypsin</td>
			<td style="width:71px;">Gibco</td>
			<td style="width:119px;">25200-056</td>
			<td>Dissociation Reagent</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">TPS 100mm Round Culture Dish</td>
			<td style="width:71px;">Midwest Scientific</td>
			<td style="width:119px;">TP93100</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Permanox 4 well slide</td>
			<td style="width:71px;">Fisher Scientific</td>
			<td style="width:119px;">1256521</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Permanox 8 well slide</td>
			<td style="width:71px;">Fisher Scientific</td>
			<td style="width:119px;">1256522</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Suction Filter Flask</td>
			<td style="width:71px;">Midwest Scientific</td>
			<td style="width:119px;">TP99500</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">15ml Conical Tube</td>
			<td style="width:71px;">Midwest Scientific</td>
			<td style="width:119px;">TP91015</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">50ml Conical Tube</td>
			<td style="width:71px;">Midwest Scientific</td>
			<td style="width:119px;">TP91050</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">2mL Round Bottom Tube</td>
			<td style="width:71px;">USA Scientific</td>
			<td style="width:119px;">1620-2700</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Small Scissor</td>
			<td style="width:71px;">FST</td>
			<td style="width:119px;">14058-11</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Small Forceps</td>
			<td style="width:71px;">FST</td>
			<td style="width:119px;">11251-20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Scalpel Handle</td>
			<td style="width:71px;">FST</td>
			<td style="width:119px;">10004-13</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">#11 Scalpel Blade</td>
			<td style="width:71px;">Roboz</td>
			<td style="width:119px;">RS-9801-11</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Non-Tissue Culture 100mm Round Petri Dish</td>
			<td style="width:71px;">Fisher Scientific</td>
			<td style="width:119px;">50-820-904</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">P1000 Pipetteman</td>
			<td style="width:71px;">Bioexpress</td>
			<td style="width:119px;">P3963-1000</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Serological Pipetteman</td>
			<td style="width:71px;">Bioexpress</td>
			<td style="width:119px;">R3073-2P</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Sterile, Non-Filtered P1000 Pipette Tips</td>
			<td style="width:71px;">Midwest Scientific</td>
			<td style="width:119px;">TD1250R</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Insulated Ice Cooler</td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Culture Hood&#160;</td>
			<td style="width:71px;">Baker</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Centrifuge&#160;</td>
			<td style="width:71px;">Eppendorf -5810R</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Hanks Balanced Salt Solution (HBSS) +/+ (w/ Ca2+, Mg2+)&#160;</td>
			<td style="width:71px;">Gibco</td>
			<td style="width:119px;">24020-117</td>
			<td>Schwannoma Culture and Media Components</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Hanks Balanced Salt Solution (HBSS) -/- (w/out Ca2+, Mg2+)</td>
			<td style="width:71px;">Gibco</td>
			<td style="width:119px;">14170-112</td>
			<td>Schwannoma Culture and Media Components</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Dulbecco&#8217;s Modified Eagle Medium (DMEM), High Glucose, w/ Phenol Red</td>
			<td style="width:71px;">Gibco</td>
			<td style="width:119px;">11965-092</td>
			<td>Schwannoma Culture and Media Components</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">N2 Supplement</td>
			<td style="width:71px;">Gibco</td>
			<td style="width:119px;">17502-048</td>
			<td>Schwannoma Culture and Media Components</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Fetal Bovine Serum</td>
			<td style="width:71px;">Gibco</td>
			<td style="width:119px;">26140-079</td>
			<td>Schwannoma Culture and Media Components</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Penicillin - Streptomycin</td>
			<td style="width:71px;">Gibco</td>
			<td style="width:119px;">15140-163</td>
			<td>Schwannoma Culture and Media Components</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Bovine Insulin (1mg/ml 200x)</td>
			<td style="width:71px;">Sigma</td>
			<td style="width:119px;">I6634</td>
			<td>Schwannoma Culture and Media Components</td>
		</tr>
	</tbody>
</table>

primary culture of human vestibular schwannomas

Vestibüler schwannomalar (VSS) tüm intrakraniyal tümörlerin% 10 taviz, Schwann hücresi vestibüler sinirin (SC) tümörler temsil eder. Vss hem TNF2 tümör baskılayıcı olarak inaktive kusurları ile ilişkili sporadik veya ailesel ya (Nörofibromatozis tip 2, NF2) formlar, meydana Gen. VSS tedavisi ancak bu tür prosedürlerin morbidite daha az invaziv tedaviler soruşturmaları tahrik olan, genellikle cerrahi rezeksiyon veya radyocerrahi olduğunu. Tarihsel olarak, taze doku örneklerinin erişim ve schwannoma hücreler immortalize değildir gerçeğinin olmaması önemli schwannoma Tümörgenez araştırılması için primer kültürlerinde kullanımını engellemiştir. Primer kültürlerin sınırlı kaynağı üstesinden gelmek için, ölümsüz kılınmış hücre kuşağı HEI193 VS HPV E6 ve E7 onkogenler ile transdüksiyonu ile üretilmiştir. Bu onkojenik transdüksiyon insan s için bir model olarak, kullanımlarını sınırlayan hücrelere, önemli moleküler ve fenotipik değişiklikler ortayachwannoma tümörler. Bu nedenle hücreler VS primer insan kültürü için basitleştirilmiş, tekrarlanabilir bir protokol göstermektedir. Bu kolayca hakim teknik daha doğru VS hastalığın karmaşıklığı özetlemek moleküler ve hücresel incelemeler için izin verir.

Doğru tümör parçalanması doku kültür tabaklarında uygun tohumlama ve oluşumun (Şekil 1) için gereklidir. Kaplama sonraki ilk günlerinde schwannoma hücrelerin tanımlanması nedeniyle hücre döküntüleri ve kırmızı kan hücrelerinin miktarda örtücü için, çoğu zaman zordur. 4 veya 5 inci inci gün, yapışık tümör parçalarının yakınında hücresi uzantıları enkaz arasında desenleri &#39;bağlama&#39; tanınabilir yaratacaktır. Daha sonraki ortam değişikliği...

Watch this Scientific Journal Video about Primary Culture of Human Vestibular Schwannomas at JoVE.com

Primary Culture of Human Vestibular Schwannomas

Vestibüler Schwannomalar (VSS) TNF2 tümör baskılayıcı gen mutasyonu ile ilişkili Schwann hücre (SC) kökenli olmayan habis tümörler vardır. Biz, moleküler ve hücresel deneysel manipülasyon ve analiz için izin verir ve insan hastalığının heterojen doğası tekrarıdır hücre kültürü VS birincil insan için yeniden üretilebilir ve etkili protokolü sunulmaktadır.

İnsan Vestibüler Schwannomalar İlköğretim Kültürü

Vestibular schwannomas (VSs) represent Schwann cell (SC) tumors of the vestibular nerve, compromising 10% of all intracranial neoplasms. VSs occur in ...

primary-culture-of-human-vestibular-schwannomas

Research

JoVE Journal

Medicine

12.7K Görüntüleme.  University of Iowa Hospitals and Clinics.  Vestibüler Schwannomalar (VSS) TNF2 tümör baskılayıcı gen mutasyonu ile ilişkili Schwann hücre (SC) kökenli olmayan habis tümörler vardır. Biz, moleküler ve hücresel deneysel manipülasyon ve analiz için izin verir ve insan hastalığının heterojen doğası tekrarıdır hücre kültürü VS birincil insan için yeniden üretilebilir ve etkili protokolü sunulmaktadır. 

Video: Primary Culture of Human Vestibular Schwannomas

Skin Punch Biopsy Explant Culture for Derivation of Primary Human Fibroblasts

Tissues and cell lines derived from an individual with disease are ideal sources to study disease-related cellular phenotypes. Patient-derived fibroblasts in this protocol have been successfully used in the derivation of induced pluripotent stem cells to model disease1. Early passages of these fibroblasts can also be used for cell-based functional assays to study specific disease pathways, mechanisms2 and subsequent drug screening approaches. The advantage of the presented protocol over enzymatic procedures are 1) the reproducibility of the technique from small amounts of tissue derived from older patients, e.g. patients affected with Parkinson's disease, 2) the technically simple approach over more challenging methodologies using enzymatic treatments, and 3) the time consideration: this protocol takes 15-20 min and can be performed immediately after biopsy arrival. Enzymatic treatments can take up to 4 hr and have the problems of overdigestion, reduction of cell viability and subsequent attachment of cells when not handled properly. This protocol describes the dissection and preparation of a 4-mm human skin biopsy for derivation of a fibroblast culture and has a very high success rate which is important when dealing with patient-derived tissue samples. In this culture, keratinocytes migrate out of the biopsy tissue within the first week after preparation. Fibroblasts appear 7-10 days after the first outgrowth of keratinocytes. DMEM high glucose media supplemented with 20% FBS favors the growth of fibroblasts over keratinocytes and fibroblasts will overgrow the keratinocytes. After 2 passages keratinocytes have been diluted out resulting in relatively homogenous fibroblast cultures which expresses the fibroblast marker SERPINH1 (HSP-47). Using this approach, 15-20 million fibroblasts can be derived in 4-8 weeks for cell banking. The skin dissection takes about 15-20 min, cells are then monitored once a day under the microscope, and media is changed every 2-3 days after attachment and outgrowth of cells.

The fibroblast explant culture protocol from human skin punch biopsies is a technically robust and simple way to derive skin cells within 4-8 weeks for banking of about 15-20 million cells at a low passage number.

İlköğretim İnsan Fibroblastlar türetilmesi için Cilt Punch Biyopsi Eksplantın Kültür

Tissues and cell lines derived from an individual with disease are ideal sources to study disease-related cellular phenotypes. Patient-derived fibroblasts ...

Tıp

In vitro Organoid Culture of Primary Mouse Colon Tumors

Several human and murine colon cancer cell lines have been established, physiologic integrity of colon tumors such as multiple cell layers, basal-apical polarity, ability to differentiate, and anoikis are not maintained in colon cancer derived cell lines. The present study demonstrates a method for culturing primary mouse colon tumor organoids adapted from Sato T et al. 1, which retains important physiologic features of colon tumors. This method consists of mouse colon tumor tissue collection, adjacent normal colon epithelium dissociation, colon tumor cells digestion into single cells, embedding colon tumor cells into matrigel, and selective culture based on the principle that tumor cells maintain growth on limiting nutrient conditions compared to normal epithelial cells.
The primary tumor organoids if isolated from genetically modified mice provide a very useful system to assess tumor autonomous function of specific genes. Moreover, the tumor organoids are amenable to genetic manipulation by virus meditated gene delivery; therefore signaling pathways involved in the colon tumorigenesis could also be extensively investigated by overexpression or knockdown. Primary tumor organoids culture provides a physiologic relevant and feasible means to study the mechanisms and therapeutic modalities for colon tumorigenesis.

In vitro Organoid Culture of Primary Mouse Colon Tumors

A simple method to establish primary murine colon tumor organoid is described. This method utilizes the feature that colon tumor cells survive and grow into organoids in media containing limited growth factors, whereas normal colon epithelial do not.

Birincil Fare Colon Tümörler in vitro organoid Kültür

Several  human and murine colon cancer cell lines have been established, physiologic  integrity of colon tumors such as multiple cell layers, basal-apical ...

Isolation, Culture, and Imaging of Human Fetal Pancreatic Cell Clusters

For almost 30 years, scientists have demonstrated that human fetal ICCs transplanted under the kidney capsule of nude mice matured into functioning endocrine cells, as evidenced by a significant increase in circulating human C-peptide following glucose stimulation1-9. However in vitro, genesis of insulin producing cells from human fetal ICCs is low10; results reminiscent of recent experiments performed with human embryonic stem cells (hESC), a renewable source of cells that hold great promise as a potential therapeutic treatment for type 1 diabetes. Like ICCs, transplantation of partially differentiated hESC generate glucose responsive, insulin producing cells, but in vitro genesis of insulin producing cells from hESC is much less robust11-17. A complete understanding of the factors that influence the growth and differentiation of endocrine precursor cells will likely require data generated from both ICCs and hESC. While a number of protocols exist to generate insulin producing cells from hESC in vitro11-22, far fewer exist for ICCs10,23,24. Part of that discrepancy likely comes from the difficulty of working with human fetal pancreas. Towards that end, we have continued to build upon existing methods to isolate fetal islets from human pancreases with gestational ages ranging from 12 to 23 weeks, grow the cells as a monolayer or in suspension, and image for cell proliferation, pancreatic markers and human hormones including glucagon and C-peptide. ICCs generated by the protocol described below result in C-peptide release after transplantation under the kidney capsule of nude mice that are similar to C-peptide levels obtained by transplantation of fresh tissue6. Although the examples presented here focus upon the pancreatic endoderm proliferation and &#946;&#160;cell genesis, the protocol can be employed to study other aspects of pancreatic development, including exocrine, ductal, and other hormone producing cells.

A protocol to isolate, culture, and image islet cell clusters (ICCs) derived from human fetal pancreatic cells is described.&#160; The method details the steps necessary to generate ICCs from tissue, culture as monolayers or in suspension as aggregates, and image for markers of proliferation and pancreatic cell fate decisions.

İnsan Fetal Pankreas Hücre Kümeleri İzolasyon, Kültür ve Görüntüleme

For almost 30 years, scientists have demonstrated that human fetal ICCs transplanted under the kidney capsule of nude mice matured into functioning ...

A Three-dimensional Tissue Culture Model to Study Primary Human Bone Marrow and its Malignancies

Tissue culture has been an invaluable tool to study many aspects of cell function, from normal development to disease. Conventional cell culture methods rely on the ability of cells either to attach to a solid substratum of a tissue culture dish or to grow in suspension in liquid medium. Multiple immortal cell lines have been created and grown using such approaches, however, these methods frequently fail when primary cells need to be grown ex vivo. Such failure has been attributed to the absence of the appropriate extracellular matrix components of the tissue microenvironment from the standard systems where tissue culture plastic is used as a surface for cell growth. Extracellular matrix is an integral component of the tissue microenvironment and its presence is crucial for the maintenance of physiological functions such as cell polarization, survival, and proliferation. Here we present a 3-dimensional tissue culture method where primary bone marrow cells are grown in extracellular matrix formulated to recapitulate the microenvironment of the human bone (rBM system). Embedded in the extracellular matrix, cells are supplied with nutrients through the medium supplemented with human plasma, thus providing a comprehensive system where cell survival and proliferation can be sustained for up to 30 days while maintaining the cellular composition of the primary tissue. Using the rBM system we have successfully grown primary bone marrow cells from normal donors and patients with amyloidosis, and various hematological malignancies. The rBM system allows for direct, in-matrix real time visualization of the cell behavior and evaluation of preclinical efficacy of novel therapeutics. Moreover, cells can be isolated from the rBM and subsequently used for in vivo transplantation, cell sorting, flow cytometry, and nucleic acid and protein analysis. Taken together, the rBM method provides a reliable system for the growth of primary bone marrow cells under physiological conditions.

In standard culture methods cells are taken out of their physiological environment and grown on the plastic surface of a dish. To study the behavior of primary human bone marrow cells we created a 3-D culture system where cells are grown under conditions recapitulating the native microenvironment of the tissue.

İlköğretim İnsan Kemik İliği ve Maligniteleri Eğitim Bir Üç boyutlu Doku Kültürü Modeli

Tissue culture has been an invaluable tool to study many aspects of cell function, from normal development to disease. Conventional cell culture methods ...

Three-dimensional Alginate-bead Culture of Human Pituitary Adenoma Cells

A three-dimensional culture method is described in which primary pituitary adenoma cells are grown in alginate beads. Alginate is a polymer derived from brown sea algae. Briefly, the tumor tissue is cut into small pieces and submitted to an enzymatic digestion with collagenase and trypsin. Next, a cell suspension is obtained. The tumor cell suspension is mixed with 1.2% sodium alginate and dropped into a CaCl2 solution, and the alginate/cell suspension is gelled on contact with the CaCl2 to form spherical beads. The cells embedded in the alginate beads are supplied with nutrients provided by the culture media enriched with 20% FBS. Three-dimensional culture in alginate beads maintains the viability of adenoma cells for long periods of time, up to four months. Moreover, the cells can be liberated from the alginate by washing the beads with sodium citrate and seeded on glass coverslips for further immunocytochemical analyses. The use of a cell culture model allows for the fixation and visualization of the actin cytoskeleton with minimal disorganization. In summary, alginate beads provide a reliable culture system for the maintenance of pituitary adenoma cells.

Three-dimensional culture in alginate beads, due to its simplicity and reproducibility, was chosen to maintain the pituitary adenoma cells. The procedures included an initial enzymatic digestion and mechanical dissociation of the tumor tissue, and the subsequent cell suspension was encapsulated in alginate beads.

İnsan Hipofiz Adenom Hücreleri üç boyutlu Aljinat-boncuk Kültür

A three-dimensional culture method is described in which primary pituitary adenoma cells are grown in alginate beads. Alginate is a polymer derived from ...

Isolation and Culture of Primary Endothelial Cells from Canine Arteries and Veins

Cardiovascular disease is studied in both human and veterinary medicine. Endothelial cells have been used extensively as an in vitro model to study vasculogenesis, (tumor) angiogenesis, and atherosclerosis. The current standard for in vitro research on human endothelial cells (ECs) is the use of Human Umbilical Vein Endothelial Cells (HUVECs) and Human Umbilical Artery Endothelial Cells (HUAECs). For canine endothelial research, only one cell line (CnAOEC) is available, which is derived from canine aortic endothelium. Although currently not completely understood, there is a difference between ECs originating from either arteries or veins. For a more direct approach to in vitro functionality studies on ECs, we describe a new method for isolating Canine Primary Endothelial Cells (CaPECs) from a variety of vessels. This technique reduces the chance of contamination with fast-growing cells such as fibroblasts and smooth muscle cells, a problem that is common in standard isolation methods such as flushing the vessel with enzymatic solutions or mincing the vessel prior to digestion of the tissue containing all cells. The technique we describe was optimized for the canine model, but can easily be utilized in other species such as human.

Novel isolation methods of primary endothelial cells from blood vessels are needed. This protocol describes a new technique that completely inverts blood vessels of interest, exposing only the endothelial side to enzymatic digestion. The resulting pure endothelial cell culture can be used to study cardiovascular diseases, disease modelling, and angiogenesis.

Köpek arter ve venlerin Primer Endotel Hücreleri İzolasyon ve Kültür

Cardiovascular disease is studied in both human and veterinary medicine. Endothelial cells have been used extensively as an in vitro model to study ...

Isolation of Primary Human Decidual Cells from the Fetal Membranes of Term Placentae

The decidua, also known as the pregnant endometrium, is a critically important reproductive tissue. Decidual cells, comprised mainly of decidualized stromal cells and immune cells, are responsible for the secretion of hormonal and inflammatory factors which are critical for successful blastocyst implantation, placental development and play a role in the initiation of labor at term and preterm. Many pregnancy complications can arise from perturbations of a fine balance of different cell populations comprising decidua. Alterations in the proportion of specific decidual cell types may disrupt these crucial processes and increase the risk of developing serious complications of pregnancy, such as embryo implantation failure, intrauterine growth restriction, preeclampsia and preterm labor. The protocol outlined here demonstrates a cost and time effective method for the isolation of primary human decidual cells collected from the fetal membranes of term placentae. By combining enzymatic digestion and gentle mechanical disruption of the decidual tissue, a high yield of decidual cells was obtained with virtually no chorion contamination. Importantly, isolated decidual cells were characterized (stromal cells (55-60%), leukocytes (35%), epithelial (1%) or trophoblast (0.01%) cells) and maintained high viability (80%) which was confirmed by multicolor imaging flow cytometry assay. This protocol is specific to the decidua parietalis and can be adapted to first and second trimester placentae. Once isolated, decidual cells can be used for a multitude of experimental applications aiming to understand the role of different decidual cell sub-populations in pregnancy complications.

This protocol demonstrates a method for the isolation of primary human decidual cells collected from the fetal membranes of term placentae which can be used for a variety of applications (i.e. immunocytochemistry, flow cytometry, etc.) aiming to study the role of different cell populations in pregnancy complications.

Dönem Placentae Fetal membranlar birincil insan Decidual hücre izolasyon

The decidua, also known as the pregnant endometrium, is a critically important reproductive tissue. Decidual cells, comprised mainly of decidualized ...

Muscle Imbalances: Testing and Training Functional Eccentric Hamstring Strength in Athletic Populations

Many hamstring injuries that occur during physical activity occur while the muscles are lengthening, during eccentric hamstring muscle actions. Opposite of these eccentric hamstring actions are concentric quadriceps actions, where the larger and likely stronger quadriceps straighten the knee. Therefore, to stabilize the lower limbs during movement, the hamstrings must eccentrically combat against the strong knee-straightening torque of the quadriceps. As such, eccentric hamstring strength expressed relative to concentric quadricep strength is commonly referred to as the &#34;functional ratio&#34; as most movements in sports require simultaneous concentric knee extension and eccentric knee flexion. To increase the strength, resiliency, and functional performance of the hamstrings, it is necessary to test and train the hamstrings at different eccentric speeds. The main purpose of this work is to provide instructions for measuring and interpreting eccentric hamstring strength. Techniques for measuring the functional ratio using isokinetic dynamometry are provided and sample data will be compared. Additionally, we briefly describe how to address hamstring strength deficiencies or unilateral strength differences using exercises that specifically focus on increasing eccentric hamstring strength.

The hamstrings are a group of muscles that are sometimes problematic for athletes, resulting in soft tissue injury in the lower limbs. To prevent such injuries, functional training of the hamstrings requires intensive eccentric contractions. Additionally, hamstring function should be tested in relation to quadricep function at different contraction speeds.

Kas dengesizlikleri: Test ve fonksiyonel eksantrik Hamstring gücü atletik nüfus eğitim

Many hamstring injuries that occur during physical activity occur while the muscles are lengthening, during eccentric hamstring muscle actions. Opposite ...

Surgical Labyrinthectomy of the Rat to Study the Vestibular System

To study the vestibular system or the vestibular compensation process, a number of methods have been developed to cause vestibular damage, including surgical or chemical labyrinthectomy and vestibular neurectomy. Surgical labyrinthectomy is a relatively simple, reliable, and rapid method. Here, we describe the surgical technique for rat labyrinthectomy. A postauricular incision is made under general anesthesia to expose the external auditory canal and the tympanic membrane, after which the tympanic membrane and the ossicles are removed without the stapes. The stapes artery, which is located between the stapes and the oval window, is a vulnerable structure and must be preserved to obtain a clear surgical field. A hole to fenestrate the vestibule is made with a 2.1-mm drill bur superior to the stapes. Then, 100% ethanol is injected through this hole and aspirated several times. Meticulous dissection under a microscope and careful bleeding control are essential to obtain reliable results. Symptoms of vestibular loss, such as nystagmus, head tilting, and a rolling motion, are seen immediately after surgery. The rotarod or rotation chair test can be used to objectively and quantitatively evaluate the vestibular function.

This protocol describes the surgical labyrinthectomy of a rat, which is a useful method for studying the vestibular system.

Vestibüler sistem çalışmaya farenin cerrahi Labyrinthectomy

To study the vestibular system or the vestibular compensation process, a number of methods have been developed to cause vestibular damage, including ...

Characterization of a Novel Human Organotypic Retinal Culture Technique

Previous human organotypic retinal culture (HORC) models have utilized detached retinas; however, without the structural support conferred by retinal pigment epithelium-choroid (RPE-choroid) and sclera, the integrity of the fragile retina can easily be compromised. The aim of this study was to develop a novel HORC model that contains the retina, RPE-choroid and sclera to maintain retinal integrity when culturing retinal explants.
After cutting circumferentially along the limbus to remove iris and lens, four deep incisions were made to flatten the eyecup. In contrast to previous HORC protocols, a trephine was used to cut through not only the retina but also the RPE-choroid and sclera. The resultant triple-layered explants were cultured for 72 h. Hematoxylin and Eosin staining (H&#38;E) was used to assess anatomical structures and retinal explants were further characterized by immunohistochemistry (IHC) for apoptosis, M&#252;ller cell integrity and retinal inflammation. To confirm the possibility of disease induction, explants were exposed to high glucose (HG) and pro-inflammatory cytokines (Cyt), to mimic diabetic retinopathy (DR). The Luminex magnetic bead assay was used to measure DR-related cytokines released into the culture medium.
H&#38;E staining revealed distinct retinal lamellae and compact nuclei in retinal explants with the underlying RPE-choroid and sclera, while retinas without the underlying structures exhibited reduced thickness and severe nuclei loss. IHC results indicated absence of apoptosis and retinal inflammation as well as preserved M&#252;ller cell integrity. The Luminex&#160;assays showed significantly increased secretion of DR-associated pro-inflammatory cytokines in retinal explants exposed to HG + Cyt relative to baseline levels at 24 h.
We successfully developed and characterized a novel HORC protocol in which retinal integrity was preserved without apoptosis or retinal inflammation. Moreover, the induced secretion of DR-associated pro-inflammatory biomarkers when exposing retinal explants to HG + Cyt suggests that this model could be used for clinically translatable retinal disease studies.

This study aims to develop a novel human organotypic retinal culture (HORC) model that prevents compromising retinal integrity during explant handling. This is achieved by culturing the retina with the overlying vitreous and the underlying retinal pigment&#160;epithelium-choroid (RPE-choroid) and sclera.

Yeni bir İnsan Organotipik Retina Kültürü Tekniğinin Karakterizasyonu

Previous human organotypic retinal culture (HORC) models have utilized detached retinas; however, without the structural support conferred by retinal ...