Generation and Characterization of Rabbit Embryonic Stem Cells

Abstract

We described the derivation of four stable pluripotent rabbit embryonic stem cell (ESC) lines, one (RF) from blastocysts fertilized in vivo and cultured in vitro and three (RP01, RP02, and RP03) from parthenogenetic blastocysts. These ESC lines have been cultivated for extended periods (RF >1 year, RP01 >8 months, RP02 >8 months, and RP03 >6 months) in vitro while maintaining expression of pluripotent ESC markers and a normal XY or XX karyotype. The ESCs from all lines expressed alkaline phosphatase, transcription factor Oct-4, stage-specific embryonic antigens (SSEA-1, SSEA-3, and SSEA-4), and the tumor-related antigens (TRA-1-60 and TRA-1-81). Similar to human and mouse ESCs, rabbit ESCs expressed pluripotency (Oct-4, Nanog, SOX2, and UTF-1) and signaling pathway genes (fibroblast growth factor, WNT, and transforming growth factor pathway). Morphologically, rabbit ESCs resembled primate ESCs, whereas their proliferation characteristics were more like those seen in mouse ESCs. Rabbit ESCs were induced to differentiate into many cell types in vitro and formed teratomas with derivatives of the three major germ layers in vivo when injected into severe combined immunodeficient mice. Our results showed that pluripotent, stable ESC lines could be derived from fertilized and parthenote-derived rabbit embryos. STEM CELLS 2007;25:481-489

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].…”

Use of pluripotent stem cells for reproductive medicine: are we there yet?

“…This pioneering achievement paved the way to isolate ESCs in various other animals such as in chicken, hamsters, cows, buffaloes, pigs, goats, horses, sheep, rabbits, dogs, cats and monkeys (rhesus and cynomolgus) (Doetschman et al 1988;Cherny et al 1994;Pain et al 1996;Li et al 2006;Mitalipov et al 2006;Brevini et al 2007;Wang et al 2007;Schneider et al 2008;Yu et al 2008;Pant & Keefer 2009;Sanna et al 2010;Kumar De et al 2011). …”

“…Although encouraging, more in-depth biomarker validation for pluripotency is still required to characterize equine ESCs as truly pluripotent (Paris & Stout 2010). Putative rabbit ESCs were first obtained from pre-implantation rabbit blastocysts (Schoonjans et al 1996), and, recently, four stable rabbit ESC lines have been derived from both in vivo produced blastocysts and parthenogenetic blastocysts (Wang et al 2007). These rabbit ESCs were morphologically similar to primate ESCs, whereas their growth rate resembled closely to mESCs (Wang et al 2007).…”