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).

이 프로토콜은 의학 스탠포드 대학의 기관 동물 보호 및 사용위원회 (IACUC)에 따라 수행 하였다. 섬 세포 복제 평가를위한 384 웰 플레이트 228 우물을 생성하기에 충분하다 - (9 주 이전 8) 남성 스프 라그 돌리 쥐, 300g - 기술 된 프로토콜은 여섯 250 섬의 격리를 위해 조정됩니다. 1. 재료 준비 <ol><li> 15cm 조직 배양 접시 (28)에서 3 일간 합류 유지 804G 래트 방광 종양 세포의 조절 매체 (RPMI...

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β 세포의 성장과 재생을 제어하는​​ 분자 경로를 연구하기위한 실험 방법은 당뇨병 연구를위한 중요한 도구입니다. 여기서, 쥐 작은 섬 기반 스크리닝 플랫폼을 제시 β 세포 복제의 작은 분자 자극을 식별하고 특성화합니다. 이 프로토콜의 대부분의 측면 쉽게 경험이 풍부한 연구자들에 의해 수행되는 동안, 몇 가지 단계가 특정 기술을 필요로한다. 첫째, 섬 격리하는 동?...

당뇨병은 포도당 항상성 교란의 공통 종점 공유 질환의 컬렉션을 포함한다. 당뇨병 아형의 병원성 메카니즘은 구별이기는하지만 감소 β 세포 질량, 즉 인슐린 생산 능력 1,2- 손실의 결과를 공유한다. 현재 당뇨병 치료 전략은 외인성 인슐린, 인슐린 생산, 인슐린 감수성 증강 약물 자극 만성 투여에 의존하고, 거의, 췌장 또는 전부를 3,4- 췌장 이식. 유감스럽게도, 이러한 전략의 성공은 짧은 수명 및 / 또는 충분히 내인성 인슐린 생산 기능을 요점을 되풀이하는 데 실패합니다. β 세포의 재생을 촉진하는 방법을 개발의 유용성에도 불구하고, 이러한 접근 방식은 존재하지 않는다. 따라서, 당뇨병의 주요 연구의 목표는 새로운 β 세포를 생성하거나 내인성 β 세포 5 질량 확장 방법을 개발하는 것이다 </sup&gt;. 이러한 배아 줄기 세포와 같은 재생 에너지 원 β - 세포 재생이 진행되고 있지만, 안전과 효율 우려가 성숙 β 세포의 확장, 우선 순위 6,7를 포함하여 다른 전략을 추구합니다. 중요한 생체 내에서 새로운 β 세포의 주된 소스는 기존의 β 세포보다는 특수 전구 세포 8,9이다. β 세포가 제한된 복제 능력 β 세포 질량의 작은 증가를 갖는 것처럼 보이지만 (~ 30 %)이 많은 당뇨 환자의 혈당 항상성을 복원하기에 충분할 수있다. 또한, β 세포 질량의 동일계 약물 자극 잠재적 인 저렴하고 확장 치료 전략이다. 여기서 β 세포의 성장을 자극하는 작은 분자를 확인하고 특성화하기위한 고급 컨텐츠 스크리닝 방법이 제공된다. 시험 관내 실험 방법의 다양한 유전자 생성물 및 / 또는 분자 그쪽을 식별하기 위해 사용될 수있다t 차 β 세포 복제를 추진하고 있습니다. β 세포 ​​복제 유도를 측정하기위한 초기의 노력은, 특정 처리 조건 (10)에 응답하여 알데히드 - 티오 닌 또는 인슐린 염색 모집단 내에서 [3 H] 티미 딘 혼입 BrdU의 혼입 또는 유사 분열 체를 측정하는 태아 설치류 pancreata 배양 또는 그대로 고립 섬 배양 물을 사용하여 11. 이 체외 접근법과 유사 검색어는 그 몇 가지 제한이 있습니다. 눈에 띄는 결함 성숙한 β 세포와 달리, 높은 기저 β 세포 복제 속도를 표시하고 명료하게 12 규제 증가하고, 태아 세포 (1)의 사용을 포함한다; (2) β 세포 복제 이벤트의 실험자에 의존 판결의 주관적 성격 (3) β 세포 복제 이벤트의 실험자에 의존하는 계산의 노동과 시간 집약적 인 특성은 실험 처리량을 지연시킨다; 핵 혼입 / 얼룩 / 외관 (4)를 사용하더라도 복제 식별TS 및 β 세포를 식별하기 위해 비 중복 세포질 염색-β 세포에 근접한 비 β 세포 복제 사건 misattribution 리드. 보다 최근 성숙한 차 β 세포가 β 세포 복제 13-16의 과발현 유전자의 영향뿐만 아니라 유전자 생성물 또는 화합물을 치료를 평가하기 위해 사용되었다. 그러나, 이들 연구는 또한 복제 이벤트, β 세포의 식별 및 / 또는 처리량을 제한하는 노동 집약적 인 단계, 예를 들어, 세포 또는 본래 췌장의 개별 슬라이드 웰 도금 세포질 staining- 또는 비특이적 방법 주관적 계수에 의존 파라핀 매립 및 처리 17. 특히, 본 명세서에서 비슷한 이미지 기반 인간의 β 세포 복제 심사 방법론은, 18 게시 된; FEA 그러나 이러한 분석의 성공적인 사용은 입증되지 않은 일차 스크리닝 인간의 아일렛의 사용은 광범위하지 않을sible. 복제 - 증진 물질을 식별하기위한 다른 전략은 β 세포주의 성장 유도를 평가하는 것이다. 초기 노력은 형질 전환 된 β 세포 라인을 같은 MIN6 세포 또는 INS 13분의 832 세포 14,19-21을 사용했다. 그러나, 이러한 세포 라인은 무제한 성장을 보여 주 잘 차별화 된 β 세포 (22)에 약간의 유사성. 따라서, 성장 유도 용량은 최소 불분명 관련성 및 요점을 되풀이하는 것이 때로는 곤란하다. 세포주 기반 스크리닝 개선 전략은 테트라 사이클린 (독시사이클린)의 부재하에 성장 체포 의존적 SV40 T 항원의 발현 23,24있는 세포 &quot;가역적으로 변환&quot;이용한다. 그러나, 이러한 세포가 독시 싸이클린 제거시 &quot;일반적인&quot;β 세포와 같은 상태로 되돌아 여부를 명확하지 않다. 불행하게도,이 세포의 사용은 즉시 유틸리티가 표시되지 않는 일반화 된 성장 촉진 화합물을 수득했다24. 전반적으로, 최소한의 자발적인 복제 작업을 표시하는 세포 유형의 성장 조절을 연구 세포 라인의 사용은 제한된 적용 가능성을 가질 수도있다. 여기에 제시된 β 세포 복제 심사 플랫폼은 가능한 한, 혈통 제한 성장 촉진 활동의 식별을 가능하게 혼합 세포 형 조성물의 섬 - 세포 배양, 멀티에게 생체 내 성장 조절에 유지하는 성숙한 기본 쥐의 β 세포를 활용 - 음 처리량 및 자동 분석을 최대화하기 위해 포맷하면 바이어스를 제거하고 처리량을 용이. 이 플랫폼의 성공적인 사용은 β 세포 복제 25, 26을 촉진 몇 가지 화합물을 식별 할 수있게되었습니다. 또한, 분석은 β 세포 복제 분자 조절 기전에 대한 통찰력을 제공하는 구조 - 활성 관계 연구 및 화학 상위 성 실험을 위해 사용되었다. 제시된 플랫폼이 성공적으로 fo를 적응했다β 세포 ​​복제 연구 렌티 바이러스의 RNAi 기반 조사 (25) 경로. 분석의 한계 (일차 전지의 사용) 확장 성을 제한 포함, 오히려 항체 기반 이미징 및 주요 섬의 사용, 사용과 관련된 (분석 인간 섬의 연구에 적용 할 수 있지만) 인간의 섬 세포, 비용보다 설치류의 사용 이미지 획득 및 분석 기능을 가진 자동 현미경의 가용성에 따라 자동화 된 이미지 획득 및 의존성을 용이하게 분산 된 섬 (파쇄 섬 구조)의. 유전자 제품이나 현장에서의 β 세포 재생을 자극하는 화합물을 식별하기위한 손쉬운 생체 검사 방법이 이상적 일 것이다하지만, 이러한 플랫폼은 아직 27 사용할 수 없습니다. 따라서, 설명 된 플랫폼은 β 세포 복제의 대부분의 측면을 조사 연구에 관심이 적합하다.

<table border="0" cellpadding="0" cellspacing="0" width="978">
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		<tr height="23">
			<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>
			<td style="width:272px;"></td>
		</tr>
		<tr height="22">
			<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>
			<td></td>
		</tr>
		<tr height="21">
			<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>
			<td></td>
		</tr>
		<tr height="21">
			<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>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">16 cm curved forceps</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">11003-16</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:364px;">12 cm curved hepostat</td>
			<td style="width:196px;">Fine Science Tools</td>
			<td style="width:147px;">13011-12</td>
			<td></td>
		</tr>
		<tr height="21">
			<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>
		</tr>
		<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>
		</tr>
		<tr height="21">
			<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>
		</tr>
		<tr height="24">
			<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>
		</tr>
		<tr height="21">
			<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<tr height="21">
			<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>
			<td></td>
		</tr>
		<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>
		</tr>
		<tr height="21">
			<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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		<tr height="21">
			<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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		<tr height="21">
			<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>
		</tr>
		<tr height="21">
			<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>
		</tr>
		<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>
		</tr>
		<tr height="21">
			<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>
		</tr>
		<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>
		</tr>
		<tr height="24">
			<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<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>
		</tr>
		<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-α 세포 또는 세포 복제를 평가하기 위해, 4 색 분석 프로토콜이 요구된다. 첫째, 객체는 DAPI 염색 (채널 1, 386 ㎚)로 식별됩니다. 다음으로, β 세포 (이벤트 1) 계산됩니다 : 개체 PDX-1 + (채널 2, 650 ㎚) 및 근교 핵 인슐린 (채널 3, 549 nm의) 협력을-표현한다. 그 후, 복제 β 세포 (이벤트 2) 계산됩니다 : (이벤트 1)이 공동 명시 기-67 (채널 4, 485 ㎚) (그림 3) 세?...

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

당뇨병 연구 분야에 중요한 도전은 섬 β 세포 복제를 조절하고 β - 세포 재생을 자극하는 방법을 개발하는 분자 메커니즘을 이해한다. 여기서 높은 콘텐츠 스크리닝 방법 식별 및 작은 분자의 β 세포의 복제를 촉진 활성을 평가하기 제시된다.

높은 콘텐츠<em&gt; 체외</em&gt; 췌장 작은 섬의 β 세포 복제 디스커버리 플랫폼

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

Murine 아일릿 절연 및 Subcapsular 신장 이식을위한 방법

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

연구

의학

High Content Screening in Neurodegenerative Diseases

Neurodegenerative 질병 높은 콘텐츠 심사

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

간 간선 재건과 부분 Orthotopic 간 이식의 쥐 모델에 대한 외과 절차

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

췌장 신경 가소성 시뮬레이션 :<em&gt; 체외</em&gt; 이중 신경 가소성 분석

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

같은 뇌 Microslices의 흥분 독성 자극<em&gt; 체외</em&gt; 스트로크의 모델

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

마우스의 자발적인 전이성 신장 세포 암 (mRCC)를 모델링에 따라 신장 절제술

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; 2 형 당뇨병의 마우스 모델 : 본래 독도 췌장 섬 세포의 분리 및 라이브 셀 2 광자 이미징

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

췌장암 새로운 치료 전략을 개발하기위한 전지 특성 줄기 공부

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

레이저 도플러: 췌 장 섬 Microvascular Vasomotion에 Vivo에서 측정 하기 위한 도구

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

로봇 중앙 Pancreatectomy 문 합 엔드 투 엔드의 응용 프로그램

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