Christensen Klaus, Roudnicky Filip, Burcin Mark, Patsch Christoph
Roche Pharmaceutical Research and Early Development, Roche Innovation Center Basel, Basel, Switzerland.
Methods Mol Biol. 2019;1994:17-29. doi: 10.1007/978-1-4939-9477-9_2.
The use of human pluripotent stem cells (hPSCs) for modeling human diseases and therapeutic applications requires differentiation methods that generate physiologically relevant cell types in a robust and standardized way. Herein, we describe an efficient and scalable monolayer protocol to convert pluripotent stem cells into vascular endothelial cells using defined culture conditions.The combinatorial use of small molecule compounds, growth factors as well as morphogens directs human pluripotent stem cells toward endothelial cells within 6 days. The protocol has the capacity to generate endothelial cells with high efficiencies of up to 80%. An additional immunomagnetic cell purification step that is based on the surface marker VE-cadherin results in a virtually pure population of endothelial cells. In a subsequent expansion step human PSC-derived endothelial cells can be further propagated, while maintaining their endothelial identity. Thus, our differentiation protocol enables the generation of hPSC-derived endothelial cells at a scale that is relevant for drug discovery campaigns or clinical applications.
利用人类多能干细胞（hPSC）进行人类疾病建模和治疗应用，需要能够以稳健且标准化的方式生成生理相关细胞类型的分化方法。在此，我们描述了一种高效且可扩展的单层培养方案，该方案利用特定的培养条件将多能干细胞转化为血管内皮细胞。小分子化合物、生长因子以及形态发生素的组合使用可在6天内将人类多能干细胞定向诱导为内皮细胞。该方案能够以高达80%的高效率生成内皮细胞。基于表面标志物VE-钙黏蛋白的额外免疫磁珠细胞纯化步骤可得到几乎纯的内皮细胞群体。在随后的扩增步骤中，人PSC来源的内皮细胞能够进一步增殖，同时保持其内皮细胞特性。因此，我们的分化方案能够以与药物研发或临床应用相关的规模生成hPSC来源的内皮细胞。
