Human-Induced Pluripotent Stem Cells Produced Under Xeno-Free Conditions
Human-Induced Pluripotent Stem Cells Produced Under Xeno-Free Conditions
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DOI：
10.1089/scd.2009.0459
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发表时间：
2010-08-01
影响因子：
4
通讯作者：
Bernardo Cibelli, Jose
中科院分区：
文献类型：
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作者：
Ross, Pablo Juan;Suhr, Steven Thomas;Bernardo Cibelli, Jose
诱导多能干细胞（iPSC）通过使从成年个体产生患者特异性多能干细胞成为可能，从根本上推进了再生医学领域。虽然细胞分化方案已经成功开发，并且人类疾病的动物模型已经证明这些细胞具有治疗由于衰老、变性、损伤和出生缺陷而产生的人类疾病和病症的潜力，但后勤问题仍然没有解决，并且阻碍了在人类临床试验中测试这些细胞的可能性。其中之一是广泛使用动物产品来产生和培养iPSC。我们在这里报告了一种无异种iPSC生成系统，该系统解决了iPSC生产的所有步骤，包括成年皮肤成纤维细胞的分离和培养，以及iPSC的生成，扩增和维持。在无异种条件下用多顺反子慢病毒载体产生的iPSC显示多能性的标志物，并产生显示3个初级胚层的指示物的胚状体（EB）。注射到裸鼠中的无异种iPSC产生经典的畸胎瘤，并且在有利于成纤维细胞的条件下培养的畸胎瘤外植体产生与输入细胞在形态上不可区分的细胞。本文描述的方案将促进临床前和临床研究中新细胞疗法的实施，可能减少监管负担，而不损害细胞的分化潜力。
Induced pluripotent stem cells (iPSCs) have radically advanced the field of regenerative medicine by making possible the production of patient-specific pluripotent stem cells from adult individuals. While cell differentiation protocols have been successfully developed, and animal models of human disease have proved that these cells have the potential to treat human diseases and conditions produced as a consequence of aging, degeneration, injury, and birth defects, logistical issues still remain unsolved and hamper the possibility of testing these cells in human clinical trials. Among them is the widely spread use of animal products for the generation and culture of iPSCs. We report here a xeno-free iPSC generation system that addresses all the steps of iPSCs production including the isolation and culture of adult skin fibroblasts, and iPSCs generation, expansion, and maintenance. iPSCs generated with a polycistronic lentiviral vector under xeno-free conditions displayed markers of pluripotency and gave rise to embryoid bodies (EBs) displaying indicators of the 3 primary germ layers. Xeno-free iPSCs injected into nude mice produced classic teratomas, and teratoma explants cultured under conditions favoring fibroblastic cells gave rise to cells morphologically indistinguishable from input cells. Protocols here described will facilitate the implementation of new cellular therapies for preclinical and clinical studies, potentially reducing the regulatory burden without compromising the differentiation potential of the cells.