Generation and differentiation of human embryonic stem cell-derived keratinocyte precursors
Generation and differentiation of human embryonic stem cell-derived keratinocyte precursors
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DOI：
10.1089/ten.2006.12.665
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发表时间：
2006-04-01
影响因子：
--
通讯作者：
Palecek, Sean P.
中科院分区：
文献类型：
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作者：
Ji, Lin;Allen-Hoffmann, B. Lynn;Palecek, Sean P.
人类胚胎干细胞（hESC）在未来的组织工程中具有巨大的潜力，有望成为几乎无限数量的所需细胞和组织类型的来源。我们已经确定了可溶性化学和细胞外基质因子，允许分离角质形成细胞前体细胞的胚胎干细胞。在明胶基质上的确定的无血清角质形成细胞生长培养基中培养由hESC形成的胚状体（EB）产生具有上皮形态的角蛋白14（K14）表达细胞。这些表达K14的细胞可以在补充有氢化可的松的培养基中传代培养，并通过添加钙诱导分层和终末分化。发现获得K14表达细胞的最佳时间用于EB形成以及EB接种后培养物的分化和生长。EB的形成是不必要的，以产生角质形成细胞前体细胞的hESC集落直接转移到角质形成细胞生长培养基允许分化成角质形成细胞谱系。随着进一步研究以优化hESC衍生的角质形成细胞前体的产生和纯化，这些细胞可以为体外或体内皮肤组织工程应用提供表皮细胞来源。
Human embryonic stem cells (hESC) hold tremendous potential in the future of tissue engineering, offering promise as a source of virtually unlimited quantities of desired cell and tissue types. We have identified soluble chemical and extracellular matrix factors that permit isolation of keratinocyte precursors from hESCs. Culturing embryoid bodies (EB) formed from hESCs in a defined serum-free keratinocyte growth medium on a gelatin matrix generated keratin 14 (K14) expressing cells with an epithelial morphology. These K14 expressing cells could be subcultured in medium supplemented with hydrocortisone and induced to stratify and terminally differentiate by addition of calcium. Optimum times for obtaining K14 expressing cells were found for EB formation and for differentiation and growth of cultures after EB plating. EB formation was not necessary to generate keratinocyte precursors; direct transfer of hESC colonies to keratinocyte growth medium permitted differentiation into the keratinocyte lineage. With further studies to optimize generation and purification of hESC-derived keratinocyte precursors, these cells could provide a source of epidermal cells for skin tissue engineering applications in vitro or in vivo.