期刊
STEM CELL RESEARCH
卷 24, 期 -, 页码 106-117出版社
ELSEVIER
DOI: 10.1016/j.scr.2017.09.001
关键词
Induced pluripotent stem cells; Rabbit; Kruppel-like factors; Transcriptome; Reprogramming; Embryo
资金
- Agence Nationale de la Recherche (ANR) [PCS-09-GENM-08, ANR-12-RPIB-0013]
- European Cooperation in Science and Technology (COST) Action [TD1101]
- HyPharm Company
- Region Rhone-Alpes [R10065CC]
- Infrastructure Nationale en Biologie et Sante INGESTEM [ANR-11-INBS-0009]
- Infrastructure Nationale en Biologie et Sante CRB-Anim [ANR-11-INBS-0003]
- IHU-B CESAME [ANR-10-IBHU-003]
- LabEx DEVweCAN [ANR-10-LABX-0061]
- LabEx CORTEX of the University of Lyon within the program 'Investissements d'Avenir' [ANR-11-LABX-0042, ANR-11-IDEX-0007]
- Agence Nationale de la Recherche (ANR) [ANR-12-RPIB-0013] Funding Source: Agence Nationale de la Recherche (ANR)
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 Kruppel-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 naive 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. (C) 2017 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
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