Name: a high-content in vitro pancreatic islet &#946;-cell replication discovery platform
Uploaded: 2016-07-16T15:00:00
Description: Watch this Scientific Journal Video about A High-content In Vitro Pancreatic Islet ÃŽÂ²-cell Replication Discovery Platform at JoVE.com

This work was supported by NIDDK grants DK098143 and DK101530 from the NIH (JPA), Stanford's Spectrum Child Health Research Institute (CHRI) and SPARK (UL1 TR001085, JPA).

Bu protokol, Stanford Üniversitesi Tıp Okulu Kurumsal Hayvan Bakım ve Kullanım Komitesi (IACUC) uygun olarak gerçekleştirilmiştir. adacık hücre replikasyonu değerlendirilmesi için 384 oyuklu plakanın 228 kuyu üretilmesi için yeterli olan - (9 haftalık 8) erkek Sprague Dawley sıçanlarında, 300 g - açıklanan protokol altı 250 doku adacığı izolasyonu ölçeklenir. 1. Malzeme Hazırlama <ol><li> 15 cm&#39;lik bir doku kültürü kabı 28 3 gün birleştiği noktada muhafaza 804G ...

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β-hücre büyümesini ve yenilenmesini kontrol moleküler yollarını inceleyerek için deneysel yöntemler diyabet araştırmacılar için önemli araçlardır. Burada, bir fare-adacık bazlı tarama platformu sunulmuştur β-hücresi çoğalmasının küçük moleküllü uyarıcıları belirlemek ve karakterize etmek. Bu protokolün en yönlerini kolaylıkla deneyimli araştırmacılar tarafından yerine getirirken, bir kaç adım belirli teknik gerektirir. İlk olarak, adacık izolasyonu...

Diyabet bozulmuş glikoz dengesinin ortak uç noktasını paylaşan bozuklukların bir koleksiyon kapsar. Diyabet alt tiplerinin patojenik mekanizma belirgin olmasına rağmen, azalmış β-hücresi kütlesi, yani insülin üretim kapasitesi 1,2 kaybı sonucu paylaşır. Halen, diyabet tedavi stratejileri eksojen insülin, insülin üretimi veya insülin duyarlılığı geliştirme farmakolojik stimülasyon kronik idaresi itimat ve nadiren, pankreas adacık ya da tüm pankreas 3,4 nakli. Ne yazık ki, bu stratejilerin başarısı kısa sürelidir ve / veya yeterince endojen insülin üretiminin işlevini özetlemek için başarısız olur. β-hücre yenilenmesini teşvik etmek için bir yöntem geliştirme programı olmasına rağmen, böyle bir yaklaşım var. Sonuç olarak, büyük bir şeker araştırma hedefi, yeni bir β-hücrelerinin üretilmesi için ya da endojen β-hücresi kütlesi 5 genişletmek için yöntemler geliştirmektir </sup&gt;. Bu embriyonik kök hücreleri gibi yenilenebilir kaynaklardan β-hücresi rejenerasyonu ilerlemektedir, ancak, güvenlik ve etkinlik ilgilidir olgun β-hücrelerinin genişlemesi, bir öncelik 6,7 dahil olmak üzere alternatif stratejileri, takip edin. Önemli olarak, in vivo olarak, yeni β-hücrelerinin baskın kaynağı önceden var olan β-hücrelerinin yerine özel bir progenitör hücreler 8,9 olan. β-hücrelerinin, sınırlı çoğaltma kapasitesine, β-hücresi kütlesindeki küçük bir artış sahip gibi görünmektedir, ancak (~% 30) pek çok diyabet hastası glukoz homeostazisin yeniden için yeterli olabilir. Ayrıca, β-hücresi kütlesi yerinde farmakolojik stimülasyon potansiyel olarak pahalı olmayan ve ölçeklenebilir bir tedavi stratejisidir. Burada β-hücresi büyümesini teşvik küçük molekülleri tanımlamak ve ayırt etmek için yüksek içerik tarama yöntemi sunulmaktadır. In vitro deney yöntemleri çeşitli gen ürünlerini ve / veya molekülleri tha tanımlamak için kullanılabilirt birincil β-hücre çoğalmasını teşvik. Β-hücre replikasyonu indüksiyonunu ölçen ilk çabalar, belirli bir tedavi koşullarında 10 yanıt olarak aldehit tiyonin veya insülin lekeli nüfus içinde [3H] timidin eklenmesini, BrdU dahil ve mitotik organları ölçmek için, fetal kemirgen pankreası kültürü ya da dokunulmamış izole adacığı kültürleri kullanılan 11. Bunlar in vitro yaklaşımlar ve yakın varyantları birkaç sınırlamaları vardır. Tanınmış eksiklikler olgun β-hücrelerinin aksine, yüksek bazal β-hücre çoğaltma oranı görüntüleyebilir ve farklı bir şekilde 12 düzenlenen büyümesi, fetal hücrelerin (1) kullanımını içerir; (2) β-hücre çoğaltma olaylarının deneyci bağımlı yargı öznel doğası; (3) β-hücre çoğaltma olaylarının deneyci bağımlı sayım emek ve zaman yoğun doğa deneysel verim geciktirir; Nükleer kuruluş / leke / görünüm (4) kullanımı bile çoğaltma tanımlamak içints ve β-hücreleri belirlemek için bir üst üste binmeyen sitoplazmik leke hücreleri olan B için yakın olmayan β-hücre replikasyonu etkinlikleri yanlış atıf yol açar. Daha yakın zamanda, olgun primer β-hücrelerinin β-hücresi çoğaltma 13-16 aşırı ifade transgenin etkisini hem de gen ürünü ya da bileşim tedavisi değerlendirmek için kullanılmıştır. Bununla birlikte, bu çalışmalar, çoğaltma etkinlikleri, β-hücresi tanımlama ve / veya hacmini sınırlayan bağımlı adımlar, örneğin, hücre ya da sağlam adacık bir kayma oyuklu kaplama için sitoplazmik staining- veya spesifik olmayan yöntemlerin ilgili sayımı dayanmıştır parafin gömme ve işleme 17. Özellikle, burada sunulan birine benzer bir görüntü tabanlı insan β-hücre çoğaltma tarama yöntemi, 18 yayımlanmıştır; FEA Ancak, bu testin başarılı bir kullanımı ortaya konmamıştır ve birinci tarama için insan adacık kullanımı genel olarak olmayabilirsible. replikasyon teşvik maddelerin belirlenmesi için alternatif bir strateji, β-hücresi çizgilerinin büyümesi indüksiyonunu değerlendirmek için. İlk çabaları dönüştürülmüş β-hücre hatları gibi min6 hücreleri ya da INS 832/13 hücreleri 14,19-21 kullanılır. Ancak, bu hücre hatları kontrolsüz büyüme gösteren ve iyi diferansiye β-hücreleri 22 az benzerlik taşımaktadır. Sonuç olarak, büyüme-indüksiyon kapasitesi, en az belirsiz alaka ve recapitulate bazen zordur. Hücre satırı tabanlı tarama için geliştirilmiş bir strateji tetrasiklin (doksisiklin) yokluğunda tutuklanan büyüme bağımlı SV40 T antijeni ifade 23,24 olan hücreler &quot;geri dönülebilir dönüştürülmüş&quot; kullanmaktadır. Ancak, bu hücrelerin doksisiklin kaldırılması üzerine &quot;normal&quot; bir β-hücre benzeri duruma dönmek belirsizdir. Ne yazık ki, bu hücrelerin kullanımı derhal kullanıma sahip görünmüyor genelleştirilmiş büyüme destekleyici bileşikler vermiştir24. Genel olarak, en az spontan replikasyon aktivitesi gösteren bir hücre tipi büyüme düzenleme çalışma hücre çizgilerinin kullanımı kısıtlı kullanımı olabilir. Burada sunulan β-hücre çoğaltma tarama platformu mümkün olduğu ölçüde, soy-kısıtlı büyüme destekleyici faaliyetlerin tanımlanmasını sağlamak için karma hücre tipi bileşiminin adacık hücre kültürleri, multi canlılarda büyüme yönetmelikte korumak için olgun birincil sıçan β-hücreleri kullanır eh verimlilik ve otomatik analiz maksimize etmek biçimlendirme önyargı ortadan kaldırmak ve verimi kolaylaştırmak için. Bu platformun başarılı kullanımı β-hücre çoğaltma 25,26 teşvik birkaç bileşiklerin belirlenmesini sağladı. Ayrıca, tahlil β-hücre çoğaltma moleküler regülasyonu içine mekanistik anlayış sağlamak için yapı-aktivite ilişkisi çalışmaları ve kimyasal epistasi deneyleri için kullanılır olmuştur. sunulan platformu başarıyla fo uyarlanmıştırβ-hücre çoğalmasının r lentiviral RNAi tabanlı soruşturma 25 yolaklarıyla. tahlilinde sınırlamaları (birincil hücre kullanımının) ölçeklenebilirlik Kısıtlı&#39;dır yerine antikor bazlı görüntüleme ve primer adacık kullanım, kullanım ile bağlantılı (deney insan doku adacığı çalışmaları için adapte edebilir) insan doku adacığı hücrelerinden, giderleri daha kemirgen kullanımı görüntü elde etme ve analiz yeteneği ile otomatik mikroskop doluluk durumuna bağlı olarak otomatik görüntü toplama ve bağımlılığını kolaylaştırmak için dağınık adacıklar (bozulmuş adacık mimarisi). Gen ürünlerinin ya da in situ β-hücresi rejenerasyonunu teşvik eden bileşiklerin belirlenmesi için basit, in vivo bir tarama yöntemi ideal olacaktır, ancak bu platform yapılmamış 27 kullanılamaz. Sonuç olarak, açıklanan platformu β-hücre çoğalmasının en yönlerini araştıran ilgilenen araştırmacı için uygundur.

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			<td height="23" style="height:23px;width:364px;">250g male Male Sprague Dawly Rat</td>
			<td style="width:196px;">Charles River</td>
			<td style="width:147px;">Stain # 400</td>
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			<td height="22" style="height:22px;width:364px;">12 cm teeth tisuue forceps</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">11021-12</td>
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			<td height="21" style="height:21px;width:364px;">11.5 cm fine scissors</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">14058-11</td>
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			<td height="21" style="height:21px;width:364px;">14.5 cm surgical scissors</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">14001-14</td>
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			<td height="21" style="height:21px;width:364px;">16 cm curved forceps</td>
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			<td style="width:147px;">11003-16</td>
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			<td height="21" style="height:21px;width:364px;">12 cm curved hepostat</td>
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			<td style="width:147px;">13011-12</td>
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			<td height="21" style="height:21px;width:364px;">12 cm scalpel handle</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">10003-12</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Tissue sieve-30 mesh</td>
			<td style="width:196px;">Bellco Glass</td>
			<td style="width:147px;">1985-85000</td>
			<td></td>
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			<td height="21" style="height:21px;width:364px;">Cizyme RI, 375,000 CDA units</td>
			<td style="width:196px;">VitaCyte</td>
			<td style="width:147px;">005-1030</td>
			<td></td>
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			<td height="24" style="height:24px;width:364px;">Hanks&#39; Balanced Salt solution (Ca++ and Mg++)</td>
			<td style="width:196px;">Gibco</td>
			<td style="width:147px;">24020-117</td>
			<td></td>
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			<td height="21" style="height:21px;width:364px;">Ketamine HCl (200 mg/20 ml)</td>
			<td style="width:196px;">JHP Pharmaceuticals</td>
			<td style="width:147px;">NDC# 42023-113-10</td>
			<td>to make anesthetic cocktail&#160;</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Xylazine (5 g/50 ml)</td>
			<td style="width:196px;">LLOYD</td>
			<td style="width:147px;">NADA# 139-236</td>
			<td>to make anesthetic cocktail&#160;</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Histopaque 1077</td>
			<td style="width:196px;">Sigma</td>
			<td style="width:147px;">H-1077</td>
			<td>to make histopaque 1100</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Histopaque 1119</td>
			<td style="width:196px;">Sigma</td>
			<td style="width:147px;">H-1119</td>
			<td>to make histopaque 1100</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Newborn Calf Serum 500 ml</td>
			<td style="width:196px;">Hyclone</td>
			<td style="width:147px;">SH30118.03</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Hanks&#39; Balanced Salt solution</td>
			<td style="width:196px;">Hyclone</td>
			<td style="width:147px;">SH30268.01</td>
			<td></td>
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			<td height="21" style="height:21px;">Dulbecco&#39;s Modified Eagle Medium/Low Glucose&#160;</td>
			<td style="width:196px;">Hyclone</td>
			<td style="width:147px;">SH30021.01</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Functionality/Viability Solution&#160;</td>
			<td style="width:196px;">Mediatech</td>
			<td style="width:147px;">99-768-CV</td>
			<td></td>
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			<td height="21" style="height:21px;width:364px;">RPMI1640 media&#160;</td>
			<td style="width:196px;">Hyclone</td>
			<td style="width:147px;">SH30096.01</td>
			<td>to make conditioned medium</td>
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			<td height="21" style="height:21px;width:364px;">804G rat bladder carcinoma cell-line</td>
			<td style="width:196px;">Available upon request</td>
			<td style="width:147px;"></td>
			<td>to make conditioned medium</td>
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			<td height="21" style="height:21px;width:364px;">Fetal Bovine Serum, Qualified</td>
			<td style="width:196px;">Gibco</td>
			<td style="width:147px;">26160</td>
			<td></td>
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			<td height="21" style="height:21px;width:364px;">GlutaMax-I</td>
			<td style="width:196px;">Gibco</td>
			<td style="width:147px;">35050-061</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Penicillin (5,000 IU/ml/Strptomycin (5 mg/ml)&#160;</td>
			<td style="width:196px;">MP Biomedicals</td>
			<td style="width:147px;">1670049</td>
			<td></td>
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			<td height="21" style="height:21px;width:364px;">Formamide 500 mL</td>
			<td style="width:196px;">Fisher BioReagents</td>
			<td style="width:147px;">BP227-500</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Antigen Unmasking Solution 250 mL (PH 6.0)</td>
			<td style="width:196px;">Vector Laboratories</td>
			<td style="width:147px;">H-3300</td>
			<td>to make 0.15 M Sodium Sitrate solution</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Dextrose, Anhydrous</td>
			<td style="width:196px;">EMD Chemicals</td>
			<td style="width:147px;">DX0145-1</td>
			<td>to make 1 M glucose solution</td>
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			<td height="24" style="height:24px;width:364px;">Nomal Donkey Serum (Powder)</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">017-000-121</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Triton X-100</td>
			<td style="width:196px;">Sigma</td>
			<td style="width:147px;">T8787-100ML</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Mouse anti-human Ki67 antibody</td>
			<td style="width:196px;">BD Biosciences</td>
			<td style="width:147px;">556003</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Goat anti-human PDX-1 antibody</td>
			<td style="width:196px;">R&#38;D Systems</td>
			<td style="width:147px;">AF2419</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Polyclonal Guinea Pig anti-insulin antibody</td>
			<td style="width:196px;">Dako</td>
			<td style="width:147px;">2016-08</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Polyclonal Rabbit anti-glucagon antibody</td>
			<td style="width:196px;">Dako</td>
			<td style="width:147px;">2014-06</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Polyclonal Rabbit anti-somatostatin antibody</td>
			<td style="width:196px;">Dako</td>
			<td style="width:147px;">2011-08</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Polyclonal chicken anti-vimentin antibody</td>
			<td style="width:196px;">abcam</td>
			<td style="width:147px;">ab24525</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Biotin-SP-conjugated, Donkey Anti-Mouse IgG</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">715-065-150</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">StreptAvidin, Alex Flour 488 conjugated&#160;</td>
			<td style="width:196px;">Invitrogen</td>
			<td style="width:147px;">S32354</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Rhodamine-conjugated Donkey Anti-Goat IgG&#160;</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">705-025-147</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Rhodamine-conjugated Donkey Anti-Guinea Pig IgG&#160;</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">706-025-148</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Rhodamine-conjugated Donkey Anti-Rabbit IgG</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">711-025-152</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Cy 5-conjugated Donkey Anti-Guinea Pig IgG&#160;</td>
			<td style="width:196px;">Jackson ImmunoResearch</td>
			<td style="width:147px;">706-175-148</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Cy 5-conjugated Donkey Anti-Goat IgG</td>
			<td>Jackson ImmunoResearch</td>
			<td>705-175-147</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Cy 5-conjugated Donkey Anti-Rabbit IgG</td>
			<td>Jackson ImmunoResearch</td>
			<td>711-175-152</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cy 5-conjugated Donkey Anti-Chicken IgG</td>
			<td>Jackson ImmunoResearch</td>
			<td style="width:147px;">703-175-155</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">DAPI</td>
			<td style="width:196px;">Millipore</td>
			<td style="width:147px;">S7113</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Disposable Reagent Reservoir 25 mL</td>
			<td style="width:196px;">Sorenson BioScience</td>
			<td style="width:147px;">39900</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">384 well, black/clear, tissue culture treated plate</td>
			<td style="width:196px;">BD Falcon</td>
			<td style="width:147px;">353962</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">96 well, black/clear, tissue culture treated plate</td>
			<td style="width:196px;">Costar</td>
			<td style="width:147px;">3603</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Multi-channel pipettor</td>
			<td style="width:196px;">Costar</td>
			<td style="width:147px;">4880</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">12-channel vaccume aspirator</td>
			<td style="width:196px;">Drummond</td>
			<td style="width:147px;">3-000-096</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Cell Scraper</td>
			<td style="width:196px;">Falcon</td>
			<td style="width:147px;">353085</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">Isotemp Water Bath Model 2223&#160;</td>
			<td style="width:196px;">Fisher Scientific</td>
			<td style="width:147px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">High-content screening instrument: ArrayScan VTI</td>
			<td style="width:196px;">Thermo Scientific</td>
			<td style="width:147px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

a high-content in vitro pancreatic islet &#946;-cell replication discovery platform

Loss of insulin-producing &#946;-cells is a central feature of diabetes. While a variety of potential replacement therapies are being explored, expansion of endogenous insulin-producing pancreatic islet &#946;-cells remains an attractive strategy. &#946;-cells have limited spontaneous regenerative activity; consequently, a crucial research effort is to develop a precise understanding of the molecular pathways that restrain &#946;-cell growth and to identify drugs capable of overcoming these restraints. Herein an automated high-content image-based primary-cell screening method to identify &#946;-cell replication-promoting small molecules is presented. Several, limitations of prior methodologies are surmounted. First, use of primary islet cells rather than an immortalized cell-line maximizes retention of in vivo growth restraints. Second, use of mixed-composition islet-cell cultures rather than a &#946;-cell-line allows identification of both lineage-restricted and general growth stimulators. Third, the technique makes practical the use of primary islets, a limiting resource, through use of a 384-well format. Fourth, detrimental experimental variability associated with erratic islet culture quality is overcome through optimization of isolation, dispersion, plating and culture parameters. Fifth, the difficulties of accurately and consistently measuring the low basal replication rate of islet endocrine-cells are surmounted with optimized immunostaining parameters, automated data acquisition and data analysis; automation simultaneously enhances throughput and limits experimenter bias. Notable limitations of this assay are the use of dispersed islet cultures which disrupts islet architecture, the use of rodent rather than human islets and the inherent limitations of throughput and cost associated with the use of primary cells. Importantly, the strategy is easily adapted for human islet replication studies. This assay is well suited for investigating the mitogenic effect of substances on &#946;-cells and the molecular mechanisms that regulate &#946;-cell growth.

β-hücre ya da α-hücresi çoğalmasını değerlendirmek için, dört renkli bir deney protokolü gereklidir. İlk olarak, nesnelerin DAPI lekelemesi (kanal 1, 386 nm) ile belirlenmiştir. Sonraki, β-hücreleri (olay 1) sayılır: nesneleri PDX-1 + (kanal 2, 650 nm) ve peri-nükleer insülin (kanal 3, 549 nm) co-ifade eder. Daha sonra, kopyalayan β-hücreleri (olay 2) sayılır: (olay 1) co-express Ki-67 (kanal 4, 485 nm) (Şekil 3)-hücreleri ß. β-hüc...

Watch this Scientific Journal Video about A High-content In Vitro Pancreatic Islet ÃŽÂ²-cell Replication Discovery Platform at JoVE.com

A High-content In Vitro Pancreatic Islet ÃƒÅ½Ã‚Â²-cell Replication Discovery Platform

diyabet araştırma alanı için kritik zorluklar adacık β-hücre çoğalmasını düzenleyen ve β-hücre yenilenmesini teşvik etmek için yöntemler geliştirmek moleküler mekanizmalarını anlamak için vardır. Burada yüksek içeriği tarama yöntemi belirlemek ve küçük moleküllerin β-hücresi replikasyon teşvik aktivitesi sunulmuştur değerlendirmek.

Bir Yüksek içerik<em&gt; İn Vitro</em&gt; Pankreas Adacık β-hücre çoğaltma Keşif Platformu

Loss of insulin-producing &#946;-cells is a central feature of diabetes. While a variety of potential replacement therapies are being explored, expansion ...

Research

JoVE Journal

Medicine

A High-content In Vitro Pancreatic Islet &#946;-cell Replication Discovery Platform

A Method for Murine Islet Isolation and Subcapsular Kidney Transplantation

Murin Islet İzolasyon ve subkapsüler Böbrek Nakli Bir Yöntem

Since the early pioneering work of Ballinger and Reckard demonstrating that transplantation of islets of Langerhans into diabetic rodents could normalize ...

High Content Screening in Neurodegenerative Diseases

Nörodejeneratif Hastalıklar Yüksek İçerik Tarama

The functional annotation of genomes, construction of molecular networks and novel drug target identification, are important challenges that need to be ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Hepatik Arter İmar Kısmi Ortotopik Karaciğer Nakli Bir Rat Modeli Cerrahi İşlemleri

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Simulating Pancreatic Neuroplasticity: In Vitro Dual-neuron Plasticity Assay

Simulating Pancreatic Neuroplasticity: In Vitro Dual-neuron Plasticity Assay

Pankreas Nöroplastisite benzetme:<em&gt; In Vitro</em&gt; Dual-nöron Plastisitesi Deneyi

Neuroplasticity is an inherent feature of the enteric nervous system and gastrointestinal (GI) innervation under pathological conditions. However, the ...

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Bir olarak Beyin Microslices arasında excitotoxic uyarılması<em&gt; In vitro</em&gt; İnme Model

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As ...

Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy

Modeling Spontaneous Metastatic Renal Cell Carcinoma mRCC in Mice Following Nephrectomy

Farelerde spontan metastatik renal hücreli karsinom (MRCC) modelleme ardından nefrektomi

One of the key challenges to improved testing of new experimental therapeutics in renal cell carcinoma (RCC) is the development of models that faithfully ...

Leprdb Mouse Model of Type 2 Diabetes: Pancreatic Islet Isolation and Live-cell 2-Photon Imaging Of Intact Islets

Leprdb Mouse Model of Type 2 Diabetes: Pancreatic Islet Isolation and Live-cell 2-Photon Imaging Of Intact Islets

Lepr<sup&gt; Db</sup&gt; Tip 2 Diyabet Fare Modeli: Bozulmamış Islets Of Pankreas Adacık İzolasyonu ve Canlı hücre 2-Foton Görüntüleme

Type 2 diabetes is a chronic disease affecting 382 million people in 2013, and is expected to rise to 592 million by 2035 1. During the past 2 decades, ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pankreas Kanseri Yeni Tedavi Stratejileri Geliştirme Hücre Özellikleri Kök incelenmesi

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo

Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo

Lazer Doppler: pankreas adacık mikrovasküler Vasomotion Vivo içinde ölçüm için bir araç

As a functional status of microcirculation, microvascular vasomotion is important for the delivery of oxygen and nutrients and the removal of carbon ...

Application of End-to-end Anastomosis in Robotic Central Pancreatectomy

Uçtan uca anastomoz Robotik Merkezi Pancreatectomy içinde uygulanması

Central pancreatectomy is carried out for the treatment of benign or low-malignant potential tumors located in the pancreatic neck or proximal part of ...