Zhang Lifei, Zheng Weiyan, Wang Yebo, Wang Yingjia, Huang He
Bone Marrow Transplantation Center, The First Affiliated Hospital, Zhejiang University School of Medicine, Zhejiang, People's Republic of China.
Cell Reprogram. 2013 Jun;15(3):216-23. doi: 10.1089/cell.2012.0064.
Human induced pluripotent stem cells (hiPSCs) need to be generated and expanded under clinically applicable culture conditions before they can be used for clinical application. In this study, we demonstrate that inactivated human mesenchymal stem cells (hMSCs) from different donors can be used as feeder cells to support the establishment and maintenance of hiPSCs. The hiPSCs we generated and expanded on hMSCs exhibited the typical morphology of human embryonic stem cells (hESCs), expressed undifferentiated pluripotent cell markers and genes, differentiated into all three germ layers via embryoid body and teratoma formation, and retained a normal chromosomal karyotype after 14 passages. However, we found that the rate of hiPSCs generation on hMSCs was 7.26%±2.09% compared with that on mouse embryonic fibroblasts (MEFs), and the calculated expansion efficiency of hiPSCs on hMSCs was lower than that on MEFs. hMSCs from various donors and different passages did not influence the results. These findings suggest that hMSCs can be used as feeder cells to derive and maintain hiPSCs, and thus provide another clinically feasible method for generating and expanding hiPSCs. However, the cytokines and adhesion molecules in this system should be identified to develop a preferable clinical culture condition for hiPSCs.
人类诱导多能干细胞（hiPSCs）在用于临床应用之前，需要在临床适用的培养条件下进行生成和扩增。在本研究中，我们证明来自不同供体的灭活人间充质干细胞（hMSCs）可用作饲养层细胞，以支持hiPSCs的建立和维持。我们在hMSCs上生成和扩增的hiPSCs表现出人类胚胎干细胞（hESCs）的典型形态，表达未分化的多能细胞标志物和基因，通过胚状体和畸胎瘤形成分化为所有三个胚层，并在传代14次后保持正常的染色体核型。然而，我们发现与在小鼠胚胎成纤维细胞（MEFs）上相比，hiPSCs在hMSCs上的生成率为7.26%±2.09%，并且计算得出的hiPSCs在hMSCs上的扩增效率低于在MEFs上的扩增效率。来自不同供体和不同传代次数的hMSCs并未影响结果。这些发现表明hMSCs可用作饲养层细胞来衍生和维持hiPSCs，从而为hiPSCs的生成和扩增提供了另一种临床可行的方法。然而，应确定该系统中的细胞因子和黏附分子，以开发更适合hiPSCs的临床培养条件。
