Feeder-free and xeno-free culture of human pluripotent stem cells using UCBS matrix.
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.
Hubei Key Laboratory of Embryonic Stem Cell Research, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, 442000, China.
Yan
Y
Ding
Human Reproductive Medical Center of Nanning Second People's Hospital, The Third Affiliated Hospital of Guangxi Medical University, Nanning, 530031, China.
Hua
H
Yang
Hubei Key Laboratory of Embryonic Stem Cell Research, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, 442000, China.
Li
L
Yu
Human Reproductive Medical Center of Nanning Second People's Hospital, The Third Affiliated Hospital of Guangxi Medical University, Nanning, 530031, China.
Chang Long
CL
Xu
College of Life Science and Bioengineering, Beijing University of Industry, Beijing, 100124, China.
Yi
Y
Zeng
Human Reproductive Medical Center of Nanning Second People's Hospital, The Third Affiliated Hospital of Guangxi Medical University, Nanning, 530031, China.
Ying
Y
Qiu
Hubei Key Laboratory of Embryonic Stem Cell Research, Taihe Hospital, Hubei University of Medicine, Shiyan, Hubei, 442000, China.
Dong Sheng
DS
Li
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D003470:Culture Media; D061252:Feeder Cells; D005260:Female; D005312:Fetal Blood / Q000502:physiology; D005346:Fibroblast Growth Factors / Q000008:administration & dosage; D005353:Fibronectins / Q000008:administration & dosage; D006801:Humans; D051379:Mice; D010446:Peptide Fragments / Q000008:administration & dosage; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology*
D003470:Culture Media; D005353:Fibronectins; D010446:Peptide Fragments; D005346:Fibroblast Growth Factors; C052888:fibroblast growth factor (1-10)
ROCK inhibitor; human pluripotent stem cells; umbilical cord blood serum