Directed Differentiation of Human-Induced Pluripotent Stem Cells to Mesenchymal Stem Cells
Directed Differentiation of Human-Induced Pluripotent Stem Cells to Mesenchymal Stem Cells
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DOI：
10.1007/978-1-4939-3584-0_17
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发表时间：
2016-01-01
期刊：
影响因子：
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通讯作者：
Tse, Hung-Fat
中科院分区：
文献类型：
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作者：
Lian, Qizhou;Zhang, Yuelin;Tse, Hung-Fat
多能基质细胞，也称为间充质干细胞（MSC），具有产生多种细胞类型的巨大潜力，包括内皮细胞、平滑肌细胞、骨、软骨和脂质细胞。该协议详细描述了如何对具有 MSC 命运的人诱导多能干细胞 (iPSC) 进行高效、谱系特异性分化。该方法采用符合临床要求的方案，采用化学成分确定的培养基、无饲养层条件以及 CD105 阳性和 CD24 阴性选择，在大约 20 天内通过分化人类多能细胞实现基于单细胞的 MSC 衍生。使用该协议生成的细胞表达典型的 MSC 表面标记，并经历类似于成人骨髓源性 MSC (BM-MSC) 的脂肪生成、成骨和软骨生成。尽管如此，与成体BM-MSC相比，iPSC-MSC表现出更高的增殖能力，高达120代，自我更新潜力没有明显丧失，并且组成型表达MSC表面抗原。使用该协议生成的 MSC 有许多应用，包括扩展到组织工程的大规模细胞数量和细胞疗法的开发。这种方法已用于挽救肢体缺血、过敏性疾病和香烟烟雾引起的肺损伤，并用于模拟哈钦森-吉尔福德早衰综合征 (HGPS) 的间充质和血管疾病。
Multipotent stromal cells, also known as mesenchymal stem cells (MSCs), possess great potential to generate a wide range of cell types including endothelial cells, smooth muscle cells, bone, cartilage, and lipid cells. This protocol describes in detail how to perform highly efficient, lineage-specific differentiation of human-induced pluripotent stem cells (iPSCs) with an MSCs fate. The approach uses a clinically compliant protocol with chemically defined media, feeder-free conditions, and a CD105 positive and CD24 negative selection to achieve a single cell-based MSCs derivation from differentiating human pluripotent cells in approximately 20 days. Cells generated with this protocol express typical MSCs surface markers and undergo adipogenesis, osteogenesis, and chondrogenesis similar to adult bone marrow-derived MSCs (BM-MSCs). Nonetheless, compared with adult BM-MSCs, iPSC-MSCs display a higher proliferative capacity, up to 120 passages, without obvious loss of self-renewal potential and constitutively express MSCs surface antigens. MSCs generated with this protocol have numerous applications, including expansion to large scale cell numbers for tissue engineering and the development of cellular therapeutics. This approach has been used to rescue limb ischemia, allergic disorders, and cigarette smoke-induced lung damage and to model mesenchymal and vascular disorders of Hutchinson-Gilford progeria syndrome (HGPS).