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

Stem Cell Research

Elen Gocza¹  Jérome Lecardonnel²  Harmonie Barasc³  Alain Pinton³  Thierry Joly⁴  Véronique Duranthon⁵  Nathalie Peynot⁵  Barbara Schmaltz-Panneau⁵  Luc Jouneau⁵  Catherine Archilla⁵  Irène Aksoy⁶  Maxime Aubry⁶  Marielle Afanassieff⁶  Lucas Medjani⁶  Ioannis Oikonomakos⁶  Chloé Mayère⁶  Pierre Osteil⁶  Yann Tapponnier⁶  Jazmine Nurse-Francis⁶  Nathalie Beaujean⁶  Anaïs Moulin⁶  Pierre Savatier⁶  Wilhelm Bouchereau⁶ 

[1] Agricultural Biotechnology Institute, H-2100 Gödöllo, Hungary;GABI, INRA, AgroParisTech, Université Paris-Saclay, 78350 Jouy-en-Josas, France;INRA, UMR 444, Génétique Cellulaire, F-31076 Toulouse, France;ISARA-Lyon, F-69007 Lyon, France;UMR BDR, INRA, ENVA, Université Paris Saclay, 78350 Jouy-en-Josas, France;Univ Lyon, Université Lyon 1, Inserm, Stem Cell and Brain Research Institute U1208, INRA USC 1361, 69500 Bron, France;

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

DOI  :  10.1016/j.scr.2017.09.001

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown