Bilousova Ganna, Roop Dennis R
Department of Dermatology, Charles C. Gates Center for Regenerative Medicine and Stem Cell Biology, University of Colorado Denver, Aurora, CO, USA.
Methods Mol Biol. 2013;961:337-50. doi: 10.1007/978-1-62703-227-8_22.
Recent advances in reprogramming somatic cells into induced pluripotent stem cells (iPSCs) offer the possibility of developing new therapeutic approaches for the treatment of a variety of diseases, including inherited skin disorders. While the ultimate goal is the use of iPSCs in the treatment of human diseases, extensive research is still required with preclinical mouse models before iPSC technology can be introduced into the clinic. Therefore, the methodology for the derivation of multipotent keratinocytes from mouse iPSCs is of particular importance since it may allow for the assessment of the feasibility of using iPSCs in the treatment of inherited skin disorders using mouse models which mimic these diseases. Here, we describe two alternative protocols for the efficient differentiation of mouse iPSCs into functional keratinocytes capable of reconstituting a normal stratified epidermis, hair follicles, and sebaceous glands when grafted onto mice. Each protocol results in a different yield and efficiency of keratinocyte derivation depending on the mouse genetic background used in the study. Both protocols employ applications of retinoic acid and bone-morphogenetic protein-4 and growth on collagen type IV-coated dishes to induce iPSC differentiation toward a keratinocyte lineage.
将体细胞重编程为诱导多能干细胞（iPSC）的最新进展为开发治疗多种疾病（包括遗传性皮肤病）的新治疗方法提供了可能性。虽然最终目标是将iPSC用于人类疾病的治疗，但在将iPSC技术引入临床之前，仍需要对临床前小鼠模型进行广泛研究。因此，从小鼠iPSC中衍生多能角质形成细胞的方法尤为重要，因为它可以通过模拟这些疾病的小鼠模型来评估使用iPSC治疗遗传性皮肤病的可行性。在此，我们描述了两种替代方案，可将小鼠iPSC高效分化为功能性角质形成细胞，当移植到小鼠身上时，这些角质形成细胞能够重建正常的分层表皮、毛囊和皮脂腺。根据研究中使用的小鼠遗传背景，每种方案产生的角质形成细胞衍生产量和效率各不相同。两种方案均采用视黄酸和骨形态发生蛋白-4，并在IV型胶原包被的培养皿上生长，以诱导iPSC向角质形成细胞谱系分化。
