Establishment of a feeder and serum-free culture system for human embryonic stem cells.

Establishment of a feeder and serum-free culture system for human embryonic stem cells. Zygote. 2020 Jan 22;:1-8 Authors: Wang L, Zhang R, Ma R, Jia G, Jian S, Zeng X, Xiong Z, Li B, Li C, Lv Z, Bai X Abstract Stem cells are an immortal cell population capable of self-renewal; they are essential for human development and ageing and are a major focus of research in regenerative medicine. Despite considerable progress in differentiation of stem cells in vitro, culture conditions require further optimization to maximize the potential for multicellular differentiation during expansion. The aim of this study was to develop a feeder-free, serum-free culture method for human embryonic stem cells (hESCs), to establish optimal conditions for hESC proliferation, and to determine the biological characteristics of the resulting hESCs. The H9 hESC line was cultured using a homemade serum-free, feeder-free culture system, and growth was observed. The expression of pluripotency proteins (OCT4, NANOG, SOX2, LIN28, SSEA-3, SSEA-4, TRA-1-60, and TRA-1-81) in hESCs was determined by immunofluorescence and western blotting. The mRNA expression levels of genes encoding nestin, brachyury and α-fetoprotein in differentiated H9 cells were determined by RT-PCR. The newly developed culture system resulted in classical hESC colonies that were round or elliptical in shape, with clear and neat boundaries. The expression of pluripotency proteins was increased, a...
Source: Zygote - Category: Reproduction Medicine Authors: Tags: Zygote Source Type: research