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Stem Cell Res. 2017 Oct;24:106-117. doi: 10.1016/j.scr.2017.09.001. Epub 2017 Sep 5.

Reprogramming of rabbit induced pluripotent stem cells toward epiblast and chimeric competency using Krüppel-like factors.

Author information

1
Univ Lyon, Université Lyon 1, Inserm, Stem Cell and Brain Research Institute U1208, INRA USC 1361, 69500 Bron, France.
2
Univ Lyon, Université Lyon 1, Inserm, Stem Cell and Brain Research Institute U1208, INRA USC 1361, 69500 Bron, France. Electronic address: marielle.afanassieff@inserm.fr.
3
ISARA-Lyon, F-69007 Lyon, France; VetAgroSup, UPSP ICE, F-69280 Marcy l'Etoile, France.
4
UMR BDR, INRA, ENVA, Université Paris Saclay, 78350 Jouy-en-Josas, France.
5
INRA, UMR 444, Génétique Cellulaire, F-31076 Toulouse, France; ENVT, F-31076 Toulouse, France.
6
GABI, INRA, AgroParisTech, Université Paris-Saclay, 78350 Jouy-en-Josas, France.
7
Agricultural Biotechnology Institute, H-2100 Gödöllo, Hungary.
8
Univ Lyon, Université Lyon 1, Inserm, Stem Cell and Brain Research Institute U1208, INRA USC 1361, 69500 Bron, France. Electronic address: pierre.savatier@inserm.fr.

Abstract

Rabbit induced pluripotent stem cells (rbiPSCs) possess the characteristic features of primed pluripotency as defined in rodents and primates. In the present study, we reprogrammed rbiPSCs using human Krüppel-like factors (KLFs) 2 and 4 and cultured them in a medium supplemented with fetal calf serum and leukemia inhibitory factor. These cells (designated rbEKA) were propagated by enzymatic dissociation for at least 30 passages, during which they maintained a normal karyotype. This new culturing protocol resulted in transcriptional and epigenetic reconfiguration, as substantiated by the expression of transcription factors and the presence of histone modifications associated with naïve pluripotency. Furthermore, microarray analysis of rbiPSCs, rbEKA cells, rabbit ICM cells, and rabbit epiblast showed that the global gene expression profile of the reprogrammed rbiPSCs was more similar to that of rabbit ICM and epiblast cells. Injection of rbEKA cells into 8-cell stage rabbit embryos resulted in extensive colonization of ICM in 9% early-blastocysts (E3.5), epiblast in 10% mid-blastocysts (E4.5), and embryonic disk in 1.4% pre-gastrulae (E6). Thus, these results indicate that KLF2 and KLF4 triggered the conversion of rbiPSCs into epiblast-like, embryo colonization-competent PSCs. Our results highlight some of the requirements to achieve bona fide chimeric competency.

KEYWORDS:

Embryo; Induced pluripotent stem cells; Krüppel-like factors; Rabbit; Reprogramming; Transcriptome

PMID:
28889080
DOI:
10.1016/j.scr.2017.09.001
[Indexed for MEDLINE]
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