Chemically fixed autologous feeder cell-derived niche for human induced pluripotent stem cell culture

Abstract

Conventional culture of human induced pluripotent (hiPS) cells requires feeder cell preparation that is time consuming and labour intensive. Alternatively, feeder-free culture of hiPS cells requires high cell seeding densities, specialized culture medium, and growth supplements, which raise the cost. Furthermore, recombinant systems require a long time for colony formation. Here, we employed chemically fixed autologous feeder cells for hiPS cell culture without additional time needed for colony formation. Treatment of (2.5% glutaraldehyde or formaldehyde for 10 min) autologous human dermal fibroblasts allowed hiPS cells to adhere and grow as undifferentiated colonies characterized by the expression of pluripotency markers such as alkaline phosphatase, Oct-3/4, and stage-specific embryonic antigen-4. Furthermore, hiPS cells cultured on chemically fixed feeders formed teratomas in vivo, characterized by all three germ layers. The chemically fixed autologous feeders may be used as a substitute for large scale culture of hiPS cells as a convenient in-house and a cost-effective method.