Zeltner Nadja, Lafaille Fabien G, Fattahi Faranak, Studer Lorenz
Developmental Biology, Center for Stem Cell Biology, Sloan-Kettering Institute for Cancer Research;
St. Giles Laboratory of Human Genetics of Infectious Diseases, The Rockefeller University.
J Vis Exp. 2014 May 22(87):51609. doi: 10.3791/51609.
Human pluripotent stem cells (hPSCs) have great potential for studying human embryonic development, for modeling human diseases in the dish and as a source of transplantable cells for regenerative applications after disease or accidents. Neural crest (NC) cells are the precursors for a large variety of adult somatic cells, such as cells from the peripheral nervous system and glia, melanocytes and mesenchymal cells. They are a valuable source of cells to study aspects of human embryonic development, including cell fate specification and migration. Further differentiation of NC progenitor cells into terminally differentiated cell types offers the possibility to model human diseases in vitro, investigate disease mechanisms and generate cells for regenerative medicine. This article presents the adaptation of a currently available in vitro differentiation protocol for the derivation of NC cells from hPSCs. This new protocol requires 18 days of differentiation, is feeder-free, easily scalable and highly reproducible among human embryonic stem cell (hESC) lines as well as human induced pluripotent stem cell (hiPSC) lines. Both old and new protocols yield NC cells of equal identity.
人类多能干细胞(hPSC)在研究人类胚胎发育、在培养皿中模拟人类疾病以及作为疾病或事故后再生应用的可移植细胞来源方面具有巨大潜力。神经嵴(NC)细胞是多种成人体细胞的前体,如来自外周神经系统和神经胶质细胞、黑素细胞和间充质细胞。它们是研究人类胚胎发育方面(包括细胞命运特化和迁移)的宝贵细胞来源。NC祖细胞进一步分化为终末分化细胞类型为体外模拟人类疾病、研究疾病机制以及为再生医学生成细胞提供了可能性。本文介绍了一种目前可用的体外分化方案的改进方法,用于从hPSC中获得NC细胞。这种新方案需要18天的分化时间,无需饲养层,易于扩展,并且在人类胚胎干细胞(hESC)系以及人类诱导多能干细胞(hiPSC)系中具有高度可重复性。新旧方案产生的NC细胞具有相同的特性。