A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells

TLDR

Human embryonic and induced pluripotent stem cells must be produced under GMP‑compliant conditions for regenerative medicine applications. The study aimed to develop a novel feeder‑free, xeno‑free culture system to overcome limitations of conventional methods. The new system, using recombinant laminin‑511 E8 and StemFit medium, supports stable, single‑cell passaging of hESCs and hiPSCs without karyotype changes and enables generation of iPSCs from fibroblasts and blood that retain differentiation potential.

Abstract

In order to apply human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) to regenerative medicine, the cells should be produced under restricted conditions conforming to GMP guidelines. Since the conventional culture system has some issues that need to be addressed to achieve this goal, we developed a novel culture system. We found that recombinant laminin-511 E8 fragments are useful matrices for maintaining hESCs and hiPSCs when used in combination with a completely xeno-free (Xf) medium, StemFit™. Using this system, hESCs and hiPSCs can be easily and stably passaged by dissociating the cells into single cells for long periods, without any karyotype abnormalities. Human iPSCs could be generated under feeder-free (Ff) and Xf culture systems from human primary fibroblasts and blood cells and they possessed differentiation abilities. These results indicate that hiPSCs can be generated and maintained under this novel Ff and Xf culture system.