Project name: Oryctolagus cuniculus

Description: Rabbit induced pluripotent stem cells (rbiPSCs) essentially display the characteristic features of primed pluripotency as defined in rodents and primates. In this 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 evidenced by the expression of transcription factors and histone marks associated with naïve pluripotency. Furthermore, microarray analysis of rbiPSCs, rbEKA cells, rabbit ICM cells, and rabbit epiblast revealed that the global gene expression profile of the reprogrammed rbiPSCs was more similar to that of rabbit ICM and epiblast cells. Injection of rbEK cells into 8-cell and 16-cell stage rabbit embryos resulted in extensive colonization of ICM in 10% mid-blastocysts (E4) 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. Overall design: In the present work, we analyzed the transcriptome of rabbit iPS cells before and after reporgramming to a naïve-like state of pluripotency. The cell lines include: (1) two independent iPS cells (referred to as B19 and B29) produced by somatic cell reprogramming with human OCT4, SOX2, c-MYC and KLF4 (described in Osteil et al. 2013 Biol Open. 2:613-628); (2) a derivative of B19, referred to as B19-EOS cells, resulting from infectioon of B19 cells with L-SIN-EOS-C(3)-EiP (EOS) lentiviral vector (Hotta et al., 2009 Nat Methods. 6:370-376); (3) two derivatives of B129-EOS, referred to as rbEKA-30 and rbEKA-9 cells, obtained by transfection of a PiggyBac vector expressing human KLF2 and KLF4. We included rabbit inner cell mass and epiblast samples as reference for naïve and primed pluripotency, respectively.Please note that the EPI and ICM samples are re-used samples from previous study [GSE79195]. However the sample titles referred in the manuscript associated with the current study are different from the previous study and correpsond to the previous titles as shown below;GSM accn. GSE79195 [PMID: 27594588] -- current studyGSM2087654 Epiblast sample 1 -- Epiblast sample 1GSM2087655 Epiblast sample 2 -- Epiblast sample 4GSM2087656 Epiblast sample 3 -- Epiblast sample 3GSM2087657 Epiblast sample 4 -- Epiblast sample 2GSM2087658 Inner Cell Mass sample 1 -- Inner Cell Mass sample 3GSM2087659 Inner Cell Mass sample 2 -- Inner Cell Mass sample 4GSM2087660 Inner Cell Mass sample 3 -- Inner Cell Mass sample 1GSM2087661 Inner Cell Mass sample 4 -- Inner Cell Mass sample 2

Data type: Transcriptome or Gene expression

Sample scope: Multiisolate

Relevance: ModelOrganism

Last updated: 2017-06-01