Stable embryonic stem cell lines in rabbits: potential small animal models for human research

Abstract

Although embryonic stem (ES) cell lines derived from mice and primates are used extensively, the development of such lines from other mammals is extremely difficult because of their rapid decline in proliferation potential and pluripotency after several passages. This study describes the establishment of rabbit ES cell lines with indefinite proliferation potential. It was found that the feeder cell density determines the fate of rabbit ES cells, and that maximum proliferation potential was obtained when they were cultured on a feeder cell density of one-sixth of the density at confluency. Higher and lower densities of feeder cells induced ES cell differentiation or division arrest. Under optimized conditions, rabbit ES cells were passaged 50 times, after which they still possessed high telomerase activity. This culture system enabled efficient gene transduction and clonal expansion from single cells. During culture, rabbit ES cells exhibited flattened monolayer cell colonies, as reported for monkey and human ES cells, and expressed pluripotency markers. Embryoid bodies and teratomas formed readily in vitro and in vivo respectively. These ES cell lines can be safely cryopreserved for later use. Thus, rabbit ES cells can be added to the list of stable mammalian ES cells, enabling the rabbit to be used as a small animal model for the study of human cell transplantation therapy.

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

“…3). Rabbit pluripotent stem cells were defined as EpiSC-type or primed state stem cells; these are rarely competent to contribute to blastocyst chimeras and are therefore developmentally and functionally distinct from naive state mouse ES cells (15,25,28).…”

“…Cells Induced by RA and SB431542-Previously, we have detected in vitro differentiated neurons and astrocytes in spontaneously differentiated rabbit pluripotent stem cells (15,25,28). Using this method, rabbit pluripotent stem cells were first induced to form EBs by suspension culture and then allowed to differentiate in the presence of FBS.…”

“…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).…”

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

“…These rabbit ESCs were morphologically similar to primate ESCs, whereas their growth rate resembled closely to mESCs (Wang et al 2007). Another study showed that the success of rabbit ESCs is determined by the feeder cell density on which they are cultured (Honda et al 2008). Rabbit ESCs reached their full pluripotency potential, similar to hESCs, when they were cultured on feeder cells with one-sixth of the density compared to the density upon confluency (Honda et al 2008).…”

“…Another study showed that the success of rabbit ESCs is determined by the feeder cell density on which they are cultured (Honda et al 2008). Rabbit ESCs reached their full pluripotency potential, similar to hESCs, when they were cultured on feeder cells with one-sixth of the density compared to the density upon confluency (Honda et al 2008). It has also been observed that rabbit ESCs rely on signalling pathways similar to hESCs to remain pluripotent, such as bFGF and Activin/Nodal signalling (Honda et al 2009).…”