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Isolation and culture of pluripotent cells from in vitro produced porcine embryos

Published online by Cambridge University Press:  24 May 2004

Ming Li
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, Guangzhou Second People's Hospital, Guangdong, and School of Life Science, Yunnan University, Yunnan, China
Yong-Hai Li
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, Guangzhou Second People's Hospital, Guangdong, and School of Life Science, Yunnan University, Yunnan, China
Yi Hou
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, Guangzhou Second People's Hospital, Guangdong, and School of Life Science, Yunnan University, Yunnan, China
Xiao-Fang Sun
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, Guangzhou Second People's Hospital, Guangdong, and School of Life Science, Yunnan University, Yunnan, China
Qingyuan Sun
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, Guangzhou Second People's Hospital, Guangdong, and School of Life Science, Yunnan University, Yunnan, China
Wei-Hua Wang
Affiliation:
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, Guangzhou Second People's Hospital, Guangdong, and School of Life Science, Yunnan University, Yunnan, China

Abstract

The present study was designed to examine whether in vitro produced porcine embryos can be used to establish an embryonic stem (ES) cell line. Porcine embryos were produced by in vitro maturation and in vitro fertilization. Embryos at the 4-cell to blastocyst stages were cultured in an ES medium containing 16% fetal bovine serum with mouse embryonic fibroblasts as a feeder layer. It was found that ES-like colonies were derived only from blastocysts. When these ES-like colonies were separated in 0.25% trypsin–0.02% EDTA solution and cultured again, ES-like colonies were further observed in the subsequent culture until the fourth passage. The cells from ES-like colonies showed positive alkaline phosphatase activity. Some cells from the colonies differentiated into several types of cells in vitro when they were cultured in the medium without feeder layers and leukemin inhibitory factor. Embryoid bodies were also formed when the cells were cultured in a suspension status. These results indicate that porcine ES-like cells can be derived from in vitro produced porcine blastocysts and these ES-like cells are pluripotent. The culture system used in the present study is useful to isolate and culture ES cells from in vitro produced porcine embryos.

Type
Research Article
Copyright
2004 Cambridge University Press

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