Derivation and characterization of ovine embryonic stem-like cell lines in semi-defined medium without feeder cells.
Domestic animal embryonic stem (ES) cells would provide an invaluable research tool for genetic breeding and the production of transgenic animals. Unfortunately, authentic domestic animals ES cells have not been established despite progress made over more than two decades. Here, we show that ovine ES-like cells can be efficiently derived and propagated in a semi-defined medium that contains N2, B27, GSK3 inhibitor (CHIR99021), and basic fibroblast growth factor (bFGF). These ovine ES-like cells had a characteristic three-dimensional appearance, showed a bFGF dose-dependence, expressed specific markers such as alkaline phosphatase (AP), Oct-4, Sox2, Nanog and can be maintained for 30 passages. Moreover, these cells differentiated in vitro into neuronal cells, and formed teratomas containing a variety of different tissues including cartilage and neural tissue when injected into kidney capsules of severe combined immunodeficiency (SCID) mice. But the cell lines fail to contribute to embryonic development upon blastocyst transplantation. To our knowledge, this is the first experiment to use semi-defined medium without feeder-cells to derive ES-like cells from ovine blastocysts, and opens the door to deriving authentic ES cells from domesticated ungulates.
The Key Lab of Animal Biotechnology of Xinjiang, Urumqi, People's Republic of China; The Key Lab of Livestock Reproduction & Biotechnology of MOA,Xinjiang Academy of Animal Science, Urumqi, People's Republic of China.
Yuncheng
Y
Zhao
Jiapeng
J
Lin
Liqin
L
Wang
Bo
B
Chen
Chuan
C
Zhou
Tong
T
Chen
Meiying
M
Guo
Sangang
S
He
Ning
N
Zhang
Chenxi
C
Liu
Mingjun
M
Liu
Juncheng
J
Huang
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D004624:Embryo Transfer; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D061252:Feeder Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D009474:Neurons / Q000166:cytology; D012756:Sheep / Q000196:embryology*