Establishment of bovine embryonic stem cells after knockdown of CDX2.
Bovine embryonic stem cells (bESCs) have not been successfully established yet. One reason could be that CDX2, as the trophectoderm regulator, expresses in bovine inner cell mass (ICM), which probably becomes a technical barrier for maintaining the pluripotency of bESCs in vitro. We hypothesized that CDX2 knockdown (CDX2-KD) could remove such negative effort, which will be helpful for capturing complete and permanent capacity of pluripotency. Expression and localization of pluripotent genes were not affected in CDX2-KD blastocysts. The CDX2-KD bESCs grew into monolayers on feeder layer. Pluripotent genes expressed at an improved levels and lasted longer time in CDX2-KD bESCs, along with down-regulation of DNA methylation on promoters of both OCT4 and SOX2. The cystic structure typical for trophoblast cells did not show during culturing CDX2-KD bESCs. CDX2-KD bESC-derived Embryoid bodies showed with compact morphology and with the improved levels of differentiations in three germ layers. CDX2-KD bESCs still carried the capacity of forming teratomas with three germ layers after long-term culture. In summary, CDX2 in bovine ICM was inducer of trophoblast lineage with negative effect on maintenance of pluripotency of bESCs. Precise regulation CDX2 expression to switch on/off will be studied next for application on establishment of bESCs.
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot 010070, China.
Xia
X
Wu
Department of Basic Medicine, Bao Tou Medical College, Bao Tou 014040, China.|Department of Pharmacy, Bao Tou Medical College, Bao Tou 014040, China.
Miao
M
Song
College of Basic Medicine, Inner Mongolia Medical University, Hohhot 010110, China.
Xi
X
Yang
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot 010070, China.
Xin
X
Liu
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot 010070, China.
Kun
K
Liu
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot 010070, China.
Cuihua
C
Jiao
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot 010070, China.
Jinze
J
Wang
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot 010070, China.
Chunling
C
Bai
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot 010070, China.
Guanghua
G
Su
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot 010070, China.
Xuefei
X
Liu
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot 010070, China.
Guangpeng
G
Li
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001755:Blastocyst / Q000166:cytology; D000071616:CDX2 Transcription Factor / Q000235:genetics* / Q000502:physiology*; D002417:Cattle; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D002467:Cell Nucleus / Q000378:metabolism; D019175:DNA Methylation; D053595:Embryonic Stem Cells / Q000166:cytology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007621:Karyotyping; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D009865:Oocytes / Q000166:cytology; D039904:Pluripotent Stem Cells / Q000166:cytology; D011401:Promoter Regions, Genetic; D055748:SOXB1 Transcription Factors / Q000378:metabolism
D000071616:CDX2 Transcription Factor; D050814:Octamer Transcription Factor-3; D055748:SOXB1 Transcription Factors; D049452:Green Fluorescent Proteins; D000469:Alkaline Phosphatase