Characterization of Embryonic Stem Cell Lines Derived from New Zealand White Rabbit Embryos

Abstract

The purposes of this study were to examine technical details in deriving and maintaining rabbit embryonic stem (rES) cell lines and to analyze their characteristics. When STO cells were used as feeder cells, no rES cell lines were established using either intact blastocysts or inner cell masses (ICMs). On the mouse embryonic fibroblasts (MEF) feeder, rES cell lines were efficiently (24%) derived. Addition of leukemia inhibitory factor (LIF) to the cells cultured on the MEF feeders further increased the derivation efficiency (57%) of rES cells. The fact that LIF induced serine-phosphorylation of STAT3 suggested LIF-dependent maintenance of rES cells. Most of the rES cell lines expressed AP, SSEA-4, Oct4, TRA-1-60, and TRA-1-81. Western blot or RT-PCR analysis also confirmed the expression of Oct4, Nanog, and Sox2. When induced to form EBs in vitro or injected to the severe combined immunodeficiency (SCID) mice, the rES cells generated embryoid bodies (EBs) and teratomas with three germ layers expressing the marker genes including MAP2, Desmin, and GATA4, respectively. In conclusion, rabbit ES cell lines can be efficiently established using our current protocols with LIF supplement. These ES cells express pluripotent stem cell markers and retain their capability to differentiate into different tissue cells. Furthermore, rES cells depend on LIF for self-renewal, likely via the JAK-STAT pathway.

Xist Deficiency and Disorders of X-Inactivation in Rabbit Embryonic Stem Cells Can Be Rescued by Transcription-Factor-Mediated Conversion

“…It is reasonable to speculate that rbESCs require maintenance under specific unknown culture conditions [5,6,8,10]. Small molecules are powerful tools for manipulating cell fate, improving derivation, and reprogramming efficiency [32][33][34], and for enabling the generation of mouse iPSCs from somatic cell types without gene insertion [35].…”

“…The rbESCs were derived as reported previously [7,8,10]. Three early passage ( p < 12) female rbESC lines (B28-6, NJ10 and NJ12) were used for this study.…”

“…The previously established rbESCs and rbiPSCs exhibited flat colony morphology and bFGF dependence and displayed many characteristics that are similar to those of hESCs [5][6][7][8][9][10][11]. It is well known that hESCs are more like mouse epiblast stem cells (mEpiSCs) than mESCs [12].…”

A study to identify and characterize the stem/progenitor cell in rabbit meniscus

“…Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as an internal control. The forward and reverse primer sequences and the resultant products were designed according to published methods, and are listed in Table 1 (Emans et al 2007;Intawicha et al 2009;Martins et al 2010;Zhao and Dong 2008). All primers were synthesized by Invitrogen (Carlsbad, CA, USA).…”

Naive-like Conversion Overcomes the Limited Differentiation Capacity of Induced Pluripotent Stem Cells

“…Rabbit models are important because the etiologies of some human diseases are more similar to those exhibited by rabbits than to those exhibited by mice. Several ES cell lines have been generated and characterized and share major important characteristics with human ES cells (23)(24)(25)(26)(27)(28). We have successfully reprogrammed liver and stomach cells of adult rabbits, which are very similar to rabbit ES cells (15).…”