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Cell Biol Int. 2015 Oct;39(10):1111-9. doi: 10.1002/cbin.10484. Epub 2015 Jun 18.

Feeder-free and xeno-free culture of human pluripotent stem cells using UCBS matrix.

Author information

1
Hubei Key Laboratory of Embryonic Stem Cell Research, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, 442000, China.
2
Human Reproductive Medical Center of Nanning Second People's Hospital, The Third Affiliated Hospital of Guangxi Medical University, Nanning, 530031, China.
3
College of Life Science and Bioengineering, Beijing University of Industry, Beijing, 100124, China.

Abstract

The ideal medium for human pluripotent stem cells (hPSCs) culture should be feeder-free, xeno-free, and completely defined. The present study aims to establish a new feeder-free and xeno-free system for culturing hPSCs. The system consists of the matrix, which was prepared from human umbilical cord blood serum (UCBS) and used to coat the culture plates, and the xeno-free medium, which was conventional serum-free hES medium supplemented with high concentrations of bFGF and Fibronectin. Compared with matrigel and mouse embryonic fibroblasts (MEFs), the UCBS matrix was proved to be equally suitable for the growth of hPSCs. After a series of experiments with different media and cytokins, the UCBS matrix was found worked the best with the basic medium (BM) supplemented with 20 ng/mL bFGF, 10 ug/mL fibronectin and Y-27632 for culture of hES cells. The hPSCs maintained normal karyotype, high proliferation rate and self-renewal ability after continuous culture more than 10 passages in this feeder-free and xeno-free system. Furthermore, a new human embryonic stem (hES) cell line was derived from discarded day 3 embryos in this newly developed culture system. In conclusion, this feeder-free and xeno-free system could not only be used to the culture hPSCs, but could also be used to derive new hES cell lines.

KEYWORDS:

ROCK inhibitor; human pluripotent stem cells; umbilical cord blood serum

PMID:
25939663
DOI:
10.1002/cbin.10484
[Indexed for MEDLINE]

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