A Novel Feeder-Free Culture System for Human Pluripotent Stem Cell Culture and Induced Pluripotent Stem Cell Derivation

Correct interactions with extracellular matrix are essential to human pluripotent stem cells (hPSC) to maintain their pluripotent self-renewal capacity during in vitro culture. hPSCs secrete laminin 511/521, one of the most important functional basement membrane components, and they can be maintained on human laminin 511 and 521 in defined culture conditions. However, large-scale production of purified or recombinant laminin 511 and 521 is difficult and expensive. Here we have tested whether a commonly available human choriocarcinoma cell line, JAR, which produces high quantities of laminins, supports the growth of undifferentiated hPSCs. We were able to maintain several human pluripotent stem cell lines on decellularized matrix produced by JAR cells using a defined culture medium. The JAR matrix also supported targeted differentiation of the cells into neuronal and hepatic directions. Importantly, we were able to derive new human induced pluripotent stem cell (hiPSC) lines on JAR matrix and show that adhesion of the early hiPSC colonies to JAR matrix is more efficient than to matrigel. In summary, JAR matrix provides a cost-effective and easy-to-prepare alternative for human pluripotent stem cell culture and differentiation. In addition, this matrix is ideal for the efficient generation of new hiPSC lines.

与细胞外基质（extracellular matrix, ECM）的正确相互作用，对于人类多能干细胞（human pluripotent stem cells, hPSCs）在体外培养过程中维持其多能自我更新能力至关重要。hPSCs可分泌层粘连蛋白（laminin）511/521——这是最重要的功能性基底膜（basement membrane）组分之一——且在限定培养体系中，可借助人源层粘连蛋白511与521维持其培养状态。然而，纯化或重组型层粘连蛋白511与521的大规模生产既困难又成本高昂。本研究测试了一株可量产层粘连蛋白的市售人绒毛膜癌细胞系JAR（human choriocarcinoma cell line JAR），是否能够支持未分化人类多能干细胞的生长。我们借助限定培养基，利用JAR细胞分泌的脱细胞基质（decellularized matrix），成功维持了多株人类多能干细胞系的培养。该JAR脱细胞基质还可支持干细胞向神经细胞与肝系细胞的定向分化。尤为重要的是，我们可在JAR基质上成功构建全新的人类诱导多能干细胞（human induced pluripotent stem cell, hiPSCs）系，并证实早期hiPSC集落与JAR基质的黏附效率优于基质胶（matrigel）。综上，JAR脱细胞基质为人类多能干细胞的培养与分化提供了一种成本低廉、制备简便的替代方案。此外，该基质还可高效用于全新hiPSC系的构建。