Chatterjee Ishita, Li Fei, Kohler Erin E, Rehman Jalees, Malik Asrar B, Wary Kishore K
Department of Pharmacology, University of Illinois at Chicago, 835 S. Wolcott Ave., Chicago, IL, 60612, USA.
Methods Mol Biol. 2016;1357:311-27. doi: 10.1007/7651_2015_203.
The study of stem cell behavior and differentiation in a developmental context is complex, time-consuming, and expensive, and for this reason, cell culture remains a method of choice for developmental and regenerative biology and mechanistic studies. Similar to ES cells, iPS cells have the ability to differentiate into endothelial cells (ECs), and the route for differentiation appears to mimic the developmental process that occurs during the formation of an embryo. Traditional EC induction methods from embryonic stem (ES) cells rely mostly on the formation of embryoid body (EB), which employs feeder or feeder-free conditions in the presence or absence of supporting cells. Similar to ES cells, iPS cells can be cultured in feeder layer or feeder-free conditions. Here, we describe the iPS cell culture methods and induction differentiation of these cells into ECs. We use anti-mouse Flk1 and anti-mouse VE-cadherin to isolate and characterize mouse ECs, because these antibodies are commercially available and their use has been described in the literature, including by our group. The ECs produced by this method have been used by our laboratory, and we have demonstrated their in vivo potential. We also discuss how iPS cells differ in their ability to differentiate into endothelial cells in culture.
在发育背景下研究干细胞行为和分化是复杂、耗时且昂贵的，因此，细胞培养仍然是发育生物学、再生生物学及机制研究的首选方法。与胚胎干细胞相似，诱导多能干细胞具有分化为内皮细胞的能力，其分化途径似乎模拟了胚胎形成过程中发生的发育过程。传统的从胚胎干细胞诱导内皮细胞的方法主要依赖于胚状体的形成，该方法在有或没有支持细胞的情况下采用饲养层或无饲养层条件。与胚胎干细胞相似，诱导多能干细胞可以在饲养层或无饲养层条件下培养。在此，我们描述诱导多能干细胞的培养方法以及将这些细胞诱导分化为内皮细胞的过程。我们使用抗小鼠Flk1和抗小鼠VE-钙黏蛋白来分离和鉴定小鼠内皮细胞，因为这些抗体有商业供应，并且其使用在文献中已有描述，包括我们小组的文献。通过这种方法产生的内皮细胞已被我们实验室使用，并且我们已经证明了它们在体内的潜能。我们还讨论了诱导多能干细胞在培养物中分化为内皮细胞的能力有何不同。
