Generation of Embryonic Stem Cell Lines from Immature Rabbit Ovarian Follicles

Abstract

In mammalian ovaries, many immature follicles remain after the dominant follicles undergo ovulation. Here we report the successful production of rabbit embryonic stem cells (ESCs) from oocytes produced by in vitro culture of immature follicles and subsequent in vitro maturation treatment. In total, we obtained 53 blastocysts from oocytes that received intracytoplasmic sperm injection followed by in vitro culture. Although only weak expression of POU5f1 was observed in the inner cell masses of in-vitro-cultured follicle-derived embryos, repeated careful cloning enabled establishment of 3 stable ESC lines. These ESC lines displayed the morphological characteristics of primed pluripotent stem cells. The ESC lines also expressed the pluripotent markers Nanog, POU5f1, and Sox2. Further, these ESCs could be differentiated into each of the 3 different germ layers both in vitro and in vivo. These results demonstrate that immature follicles from rabbits can be used to generate ESCs. Moreover, the use of rabbit oocytes as a cell source provides an experimental system that closely matches human reproductive and stem cell physiology.

Derivation of Rabbit Embryonic Stem Cells from Vitrified–Thawed Embryos

“…A limited number of groups have worked to derive rabbit PSCs (Fang et al, 2006;Graves and Moreadith, 1993;Honda et al, 2008;Honda et al, 2013;Intawicha et al, 2009;Jiang et al, 2014;Osteil et al, 2013;Tancos et al, 2012;Teramura et al, 2013;Wang et al, 2007;Wang et al, 2008;Xue et al, 2012). Capturing the ground state (interchangeably referred to as ''authentic,'' ''naïve,'' and ''germ line transmitting'' in this article) of rbESCs has been a challenge.…”

Sexual Determination and Differentiation During Embryonic and Fetal Development of New Zealand Rabbit Females

“…Fang et al (2006) studied different cell lines derived from in vitro fertilization, parthenogenesis and somatic cells with nuclear transfer seeking to reach different cell lines and their behaviors for cellular therapies. Teramura et al (2013) sought a new source of embryonic stem cells derived from immature primordial rabbit follicles for new treatments. Another paper that used rabbit as an animal model was by Abd-Allah et al (2013), who chemically induced the formation of ovarian failure and then treated it with mesenchymal stem cells and thus observed[HML1] the pathway of these cells in this disease.…”