Characterization of embryonic stem cell lines derived from New Zealand white rabbit embryos.
The purposes of this study were to examine technical details in deriving and maintaining rabbit embryonic stem (rES) cell lines and to analyze their characteristics. When STO cells were used as feeder cells, no rES cell lines were established using either intact blastocysts or inner cell masses (ICMs). On the mouse embryonic fibroblasts (MEF) feeder, rES cell lines were efficiently (24%) derived. Addition of leukemia inhibitory factor (LIF) to the cells cultured on the MEF feeders further increased the derivation efficiency (57%) of rES cells. The fact that LIF induced serine-phosphorylation of STAT3 suggested LIF-dependent maintenance of rES cells. Most of the rES cell lines expressed AP, SSEA-4, Oct4, TRA-1-60, and TRA-1-81. Western blot or RT-PCR analysis also confirmed the expression of Oct4, Nanog, and Sox2. When induced to form EBs in vitro or injected to the severe combined immunodeficiency (SCID) mice, the rES cells generated embryoid bodies (EBs) and teratomas with three germ layers expressing the marker genes including MAP2, Desmin, and GATA4, respectively. In conclusion, rabbit ES cell lines can be efficiently established using our current protocols with LIF supplement. These ES cells express pluripotent stem cell markers and retain their capability to differentiate into different tissue cells. Furthermore, rES cells depend on LIF for self-renewal, likely via the JAK-STAT pathway.
Intawicha
Payungsuk
P
Department of Animal Science, National Chung Hsing University, Taichung, Taiwan, Republic of China.
Ou
Yao-Wen
YW
Lo
Neng-Wen
NW
Zhang
Su-Chun
SC
Chen
Yin-Zhi
YZ
Lin
Tzu-An
TA
Su
Hong-Lin
HL
Guu
Hwa-Fen
HF
Chen
Ming-Jer
MJ
Lee
Kun-Hsiung
KH
Chiu
Yung-Tsung
YT
Ju
Jyh-Cherng
JC
D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D003893:Desmin / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D005347:Fibroblasts / Q000378:metabolism; D050991:GATA4 Transcription Factor / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology; D051379:Mice; D016513:Mice, SCID; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism; D011817:Rabbits; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D055746:Stage-Specific Embryonic Antigens / Q000378:metabolism; D013724:Teratoma / Q000378:metabolism*
D003893:Desmin; D050991:GATA4 Transcription Factor; D018398:Homeodomain Proteins; D053605:Leukemia Inhibitory Factor; D008869:Microtubule-Associated Proteins; D050814:Octamer Transcription Factor-3; D011509:Proteoglycans; D055748:SOXB1 Transcription Factors; D055746:Stage-Specific Embryonic Antigens; C066584:stage-specific embryonic antigen-4