Department of Neurology, University of Michigan, Ann Arbor, MI 48109, USA; The Human Stem Cell and Gene Editing Core, University of Michigan, Ann Arbor, MI 48109, USA.
Department of Neurology, University of Michigan, Ann Arbor, MI 48109, USA.
STAR Protoc. 2023 Dec 15;4(4):102629. doi: 10.1016/j.xpro.2023.102629. Epub 2023 Oct 9.
Single-cell clonal selection is a critical procedure for generating a homogeneous population of human pluripotent stem cells. Here, we present a protocol that repurposes the STRIPPER Micropipetter, normally used for in vitro fertilization, to pick single stem cells. We describe steps for tool and reagent preparation, single-cell picking, and colony passaging. We then detail procedures for amplification and analysis. Our protocol does not require cell sorting and produces homogenous clonal cultures with more than 50% survival rate. For complete details on the use and execution of this protocol, please refer to Deng et al..
单细胞克隆选择是生成同质的人类多能干细胞群体的关键步骤。在这里，我们介绍了一种方案，该方案重新利用了通常用于体外受精的 STRIPPER Micropipetter 来挑选单个干细胞。我们描述了工具和试剂准备、单细胞挑选和集落传代的步骤。然后，我们详细介绍了扩增和分析的程序。我们的方案不需要细胞分选，并且产生具有超过 50%存活率的同质克隆培养物。有关使用和执行此方案的完整详细信息，请参阅 Deng 等人的研究。
