Generation of naive-like porcine-induced pluripotent stem cells capable of contributing to embryonic and fetal development.
In pluripotent stem cells (PSCs), there are 2 types: naive and primed. Only the naive type has the capacity for producing chimeric offspring. Mouse PSCs are naive, but human PSCs are in the primed state. Previously reported porcine PSCs appear in the primed state. In this study, putative naive porcine-induced pluripotent stem cells (iPSCs) were generated. Porcine embryonic fibroblasts were transduced with retroviral vectors expressing Yamanaka's 4 genes. Emergent colonies were propagated in the presence of porcine leukemia inhibitory factor (pLIF) and forskolin. The cells expressed pluripotency markers and formed embryoid bodies, which gave rise to cell types from all 3 embryonic germ layers. The naive state of the cells was demonstrated by pLIF dependency, 2 active X chromosomes (when female), absent MHC class I expression, and characteristic gene expression profiles. The porcine iPSCs contributed to the in vitro embryonic development (11/24, 45.8%) as assessed by fluorescent markers. They also contributed to the in utero fetal development (11/71, 15.5% at day 23; 1/13, 7.7% at day 65). This is the first demonstration of macroscopic fluorescent chimeras derived from naive-like porcine PSCs, although adult chimeras remain to be produced.
Division of Regenerative Medicine, Center for Molecular Medicine, Jichi Medical University, Tochigi, Japan.
Shuh-hei
SH
Fujishiro
Kazuaki
K
Nakano
Yoshihisa
Y
Mizukami
Takuya
T
Azami
Yoshikazu
Y
Arai
Hitomi
H
Matsunari
Rikiya
R
Ishino
Takashi
T
Nishimura
Masahito
M
Watanabe
Tomoyuki
T
Abe
Yutaka
Y
Furukawa
Kazuhiro
K
Umeyama
Shinya
S
Yamanaka
Masatsugu
M
Ema
Hiroshi
H
Nagashima
Yutaka
Y
Hanazono
D000818:Animals; D030801:Animals, Genetically Modified; D053624:Blastocyst Inner Cell Mass / Q000166:cytology*; D002478:Cells, Cultured; D002678:Chimera; D019175:DNA Methylation; D058732:Embryoid Bodies / Q000378:metabolism; D047108:Embryonic Development; D005260:Female; D047109:Fetal Development; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism* / Q000637:transplantation; D009028:Morula / Q000166:cytology; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D034421:Sus scrofa; D014161:Transduction, Genetic; D019076:Transgenes; D014960:X Chromosome / Q000235:genetics
D050814:Octamer Transcription Factor-3