Development of a simple, repeatable, and cost-effective extracellular matrix for long-term xeno-free and feeder-free self-renewal of human pluripotent stem cells

作者：Pakzad Mohammad; Ashtiani Mohammad Kazemi; Mousavi Gargari Seyed Latif; Baharvand Hossein^*

来源：Histochemistry and Cell Biology, 2013, 140(6): 635-648.

DOI：10.1007/s00418-013-1144-3

• Conditioned medium
• Extracellular matrix
• Human-induced pluripotent stem cells
• Human embryonic stem cells
• Self-renewal

Given the potential importance of human pluripotent stem cells (hPSCs) in translational research and regenerative medicine, the aim of the present study was to develop a simple, safe, and cost-effective substrate to expand hPSCs. We report the development of an extracellular matrix (ECM), designated "RoGel," based on conditioned medium (CM) of human fibroblasts under serum- and xeno-free culture conditions. The long-term self-renewal of hPSCs on RoGel was also assessed. The results showed that self-renewal, pluripotency, plating efficiency, and cloning efficiency of hPSCs on this newly developed ECM were similar to those of Matrigel, the conventional mouse-cell line-derived ECM. The cells had the capability to passage mechanically on a cold surface, which resulted in their long-term maintenance with normal karyotype. We have demonstrated that CM-coated plates preserved for 1 year at room temperature maintained the capability of hPSC expansion. This ECM provides an attractive hPSC culture platform for both research and future therapeutic applications.

• 出版日期2013-12