Abstract
Rabbits are commonly used as laboratory animal models to investigate human diseases and phylogenetic development. However, pluripotent stem cells that contribute to germline transmission have yet to be established in rabbits. The transcription factor Oct4, also known as Pou5f1, is considered essential for the maintenance of the pluripotency of stem cells. Hence, pluripotent cells can be identified by monitoring Oct4 expression using a well-established Oct4 promoter-based reporter system. This study developed a rabbit Oct4 promoter-based enhanced green fluorescent protein (EGFP) reporter system by transfecting pROP2-EGFP into rabbit fetal fibroblasts (RFFs). The transgenic RFFs were used as donor cells for somatic cell nuclear transfer (SCNT). The EGFP expression was detected in the blastocysts and genital ridges of SCNT fetuses. Fibroblasts and neural stem cells (NSCs) were derived from the SCNT fetuses. EGFP was also reactivated in blastocysts after the second SCNT, and induced pluripotent stem cells (iPSCs) were obtained after reprogramming using Yamanaka's factors. The results above indicated that a rabbit reporter system used to monitor the differentiating status of cells was successfully developed.
Highlights
Embryon ic stem cells (ESCs) are unique cell populations with self-renewal, proliferating and pluripotent properties
We used the rabbit ESCs (Rb-ESCs) to test the specificity of the pROP2EGFP vector by cell transfection and found that enhanced green fluorescent protein (EGFP) fluorescence was partly expressed in the colonies (Fig. 1B)
Oct4 is considered as an essential marker used to monitor pluripotent stem cells (e.g., ESCs, Induced pluripotent stem cells (iPSCs) and primordial germ cells (PGCs))
Summary
Embryon ic stem cells (ESCs) are unique cell populations with self-renewal, proliferating and pluripotent properties. ESCs have been used to study developmental biology, genetic modification and regenerative medicine. Induced pluripotent stem cells (iPSCs) have been generated from several mammals and opened a new frontier in regenerative medicine [1,2,3,4,5]. Before ESCs or iPSCs are clinically applied to humans, extensive pre-clinical studies should be performed using suitable animal models to assess the safety, efficacy and long-term survival of these cells. Surgical operations can be performed more with rabbits (Oryctolagus cuniculus) as laboratory animal models than mice or rats because of rabbits’ large size. The physiology of rabbits is close to that of primates
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