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Isolation and culture of embryonic stem cells from porcine blastocysts
Author(s) -
Li Ming,
Zhang Dong,
Hou Yi,
Jiao Lihong,
Zheng Xing,
Wang WeiHua
Publication year - 2003
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.10301
Subject(s) - biology , inner cell mass , embryonic stem cell , blastocyst , embryoid body , cell culture , microbiology and biotechnology , embryo , alkaline phosphatase , stem cell , cell , induced pluripotent stem cell , andrology , embryogenesis , anatomy , enzyme , genetics , biochemistry , gene , medicine
This study was conducted to establish embryonic stem (ES) cell lines from porcine blastocysts. Blastocysts were collected from China miniature pigs at day 7–9 of pregnancy. Embryos were either directly (intact embryos) cultured on mitomysin C‐inactivated murine embryonic fibroblasts (MEF) as feeder layers, or were used to isolate the inner cell masses (ICM) by enzyme digestive method and then cultured. It was found that enzyme digestive method could isolate ICMs without any damages of cells in all blastocysts (28). All ICMs attached to the feeder layers. Primary cell colonies were formed in 68% of ICM culture and 28% of intact blastocyst culture. Two ES cell lines derived from ICM passed six subcultures (passages). These cells morphologically resembled mouse ES cells and consistently expressed alkaline phosphatase activity. When the ES cells were cultured in a medium without feeder layer and leukemin inhibitory factor, they differentiated into several types of cells including neuron‐like, smooth muscle‐like, and epithelium‐like cells. Some cells formed embryoid bodies in a suspension culture. These results indicate that porcine ES cell line can be established under the present experimental conditions and these ES cells are pluripotent. Mol. Reprod. Dev. 65: 429–434, 2003. © 2003 Wiley‐Liss, Inc.