An optimized protocol for isolation of murine pancreatic single cells with high yield and purity.
An optimized protocol for isolation of murine pancreatic single cells with high yield and purity.
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一种优化的方案，用于分离具有高产量和纯度的鼠胰腺单细胞。
DOI：
10.1016/j.xpro.2024.102836
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发表时间：
2024-03-15
期刊：
影响因子：
--
通讯作者：
Wang, Timothy C.
中科院分区：
文献类型：
--
作者：
Wu, Feijing;Jiang, Zhengyu;Qian, Jin;Kobayashi, Hiroki;Waterbury, Quin T.;White, Ruth A.;Ochiai, Yosuke;Zhi, Xiaofei;Tu, Ruhong;Zheng, Biyun;Shi, Qiongyu;Zamechek, Leah B.;Wang, Timothy C.
在这里，我们提出了一个协议，用于快速分离单细胞从小鼠胰腺，最大限度地减少外分泌细胞中的消化酶造成的损害。我们将指导您完成优化切割顺序、酶组成和操作程序的步骤，从而获得高产量的活胰腺单细胞。该方案可应用于广泛的研究领域，包括单细胞测序、基因表达谱、原代细胞培养，甚至球状体或类器官的发育。有关本方案使用和执行的完整详细信息，请参见Jiang等人。（2023年）。分离胰腺组织，同时最大限度地减少酶损伤进行有效的胰腺组织消化程序使用流式细胞术标记和分离特定的细胞类型培养和分析分离的胰腺单细胞出版商说明：进行任何实验方案都需要遵守当地机构的实验室安全和伦理指南。在这里，我们提出了一个协议，用于快速分离单细胞从小鼠胰腺，最大限度地减少外分泌细胞中的消化酶造成的损害。我们将指导您完成优化切割顺序、酶组成和操作程序的步骤，从而获得高产量的活胰腺单细胞。该方案可以应用于广泛的研究领域，包括单细胞测序，基因表达谱，原代细胞培养，甚至球体或类器官的发育。
Here, we present a protocol for rapidly isolating single cells from the mouse pancreas, minimizing damage caused by digestive enzymes in exocrine cells. We guide you through steps to optimize the dissection sequence, enzyme composition, and operational procedures, resulting in high yields of viable pancreatic single cells. This protocol can be applied across a wide range of research areas, including single-cell sequencing, gene expression profiling, primary cell culture, and even the development of spheroids or organoids. For complete details on the use and execution of this protocol, please refer to Jiang et al. (2023). Isolate pancreatic tissue while minimizing enzyme damage Conduct an efficient pancreatic tissue digestion procedure Label and isolate specific cell types using flow cytometry Culture and analyze isolated pancreatic single cells Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for rapidly isolating single cells from the mouse pancreas, minimizing damage caused by digestive enzymes in exocrine cells. We guide you through steps to optimize the dissection sequence, enzyme composition, and operational procedures, resulting in high yields of viable pancreatic single cells. This protocol can be applied across a wide range of research areas, including single-cell sequencing, gene expression profiling, primary cell culture, and even the development of spheroids or organoids.
影响因子：
24.5
作者：
Takahashi, Ryota;Macchini, Marina;Wang, Timothy C.
通讯作者：
Wang, Timothy C.
影响因子：
23.9
作者：
Jiang，Zhengyu;Wu，Feijing;Wang，Timothy C.
通讯作者：
Wang，Timothy C.
影响因子：
64.5
作者：
Boj SF;Hwang CI;Baker LA;Chio II;Engle DD;Corbo V;Jager M;Ponz-Sarvise M;Tiriac H;Spector MS;Gracanin A;Oni T;Yu KH;van Boxtel R;Huch M;Rivera KD;Wilson JP;Feigin ME;Öhlund D;Handly-Santana A;Ardito-Abraham CM;Ludwig M;Elyada E;Alagesan B;Biffi G;Yordanov GN;Delcuze B;Creighton B;Wright K;Park Y;Morsink FH;Molenaar IQ;Borel Rinkes IH;Cuppen E;Hao Y;Jin Y;Nijman IJ;Iacobuzio-Donahue C;Leach SD;Pappin DJ;Hammell M;Klimstra DS;Basturk O;Hruban RH;Offerhaus GJ;Vries RG;Clevers H;Tuveson DA
通讯作者：
Tuveson DA