[Different types of feeder cells for maintenance of human embryonic stem cells].
OBJECTIVE
To compare the effects of different types of feeder cells on supporting undifferentiation and high proliferation of human embryonic stem cells (hESC).


METHODS
hESC were seeded on mouse embryonic fibroblasts (MEF), human marrow stromal cells (hMSC), and human foreskin fibroblasts (hFF), respectively. Colony number, cell quantity after digestion, and survival rate were observed by alkaline phosphatase (AP) staining and Trypan blue, and the biological properties of hESC after 5 passages were observed by immunofluorescence staining.


RESULTS
Although all the three feeder layers could support the formation of hESC colonies and maintain pluripotency, the morphology of colonies on different feeder layers remarkably varied. The stage-specific embryonic antigen-3 and AP staining were positive on three types of feeders. The number of colonies, number of cells produced, and cell survival rates were significantly higher on MEF than on human feeder cells (P < 0.01). Furthermore, the number of AP-positive colonies and cell quantity were also significantly higher on hMSC than on hFF (P < 0.01).


CONCLUSIONS
All three types of feeder cells are able to support the growth of hMSC, although MEF are more favourable for the proliferation. Two types of human feeder cells lay the foundation for the removal of animal-derived hESC culture system. hMSC is superior to hFF in supporting the proliferation of hESC.
Zhu
Wan-Wan
WW
Department of Cell Therapy, Beijing Institute of Geriatrics, Xuanwu Hospital, Capital Medical University, Beijing 100053, China.
Li
Ning
N
Wang
Fang
F
Fu
Lin-Lin
LL
Xu
Yan-Ling
YL
Guan
Yun-Qian
YQ
Zhang
Yu
Y
D000818:Animals; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000378:metabolism; D018920:Coculture Techniques / Q000379:methods*; D053595:Embryonic Stem Cells; D061252:Feeder Cells; D005347:Fibroblasts; D006801:Humans; D051379:Mice; D055746:Stage-Specific Embryonic Antigens / Q000378:metabolism
D015295:Antigens, Tumor-Associated, Carbohydrate; D055746:Stage-Specific Embryonic Antigens; C052924:stage-specific embryonic antigen-3