Insights into Species Preservation: Cryobanking of Rabbit Somatic and Pluripotent Stem Cells

Abstract

Induced pluripotent stem cells (iPSCs) are obtained by genetically reprogramming adult somatic cells via the overexpression of specific pluripotent genes. The resulting cells possess the same differentiation properties as blastocyst-stage embryonic stem cells (ESCs) and can be used to produce new individuals by embryonic complementation, nuclear transfer cloning, or in vitro fertilization after differentiation into male or female gametes. Therefore, iPSCs are highly valuable for preserving biodiversity and, together with somatic cells, can enlarge the pool of reproductive samples for cryobanking. In this study, we subjected rabbit iPSCs (rbiPSCs) and rabbit ear tissues to several cryopreservation conditions with the aim of defining safe and non-toxic slow-freezing protocols. We compared a commercial synthetic medium (STEM ALPHA.CRYO3) with a biological medium based on fetal bovine serum (FBS) together with low (0–5%) and high (10%) concentrations of dimethyl sulfoxide (DMSO). Our data demonstrated the efficacy of a CRYO3-based medium containing 4% DMSO for the cryopreservation of skin tissues and rbiPSCs. Specifically, this medium provided similar or even better biological results than the commonly used freezing medium composed of FBS and 10% DMSO. The results of this study therefore represent an encouraging first step towards the use of iPSCs for species preservation.

Molecular phenotyping uncovers differences in basic housekeeping functions among closely related species of hares (Lepus spp., Lagomorpha: Leporidae)

“…For this reason, significant efforts have been made to obtain fibroblast‐derived iPSCs from vulnerable and endangered species (Ben‐Nun et al, 2011; Ramaswamy et al, 2015; Verma et al, 2013; Weeratunga et al, 2020). Successful generation of iPSCs from skin derived fibroblasts has been described in rabbits (Gavin‐Plagne et al, 2020), but the method has not been validated for hares.…”

Pluripotent Stem Cells for Transgenesis in the Rabbit: A Utopia?

“…In any case, rbPSCs remain valuable tools for transgenesis, as they can be genetically modified in vitro and especially verified for mutations created before producing the transgenic rabbit strains. Moreover, iPSC technology is also of interest for the preservation of species biodiversity [151], by allowing easy sampling from valuable animals without any breeding steps. This would consequently enlarge the number of frozen samples with somatic tissues.…”

Creating Ex Situ Protected Bioreservoirs as a Powerful Strategy for the Reproductive Biotechnology-Mediated Rescue of Threatened Polish Livestock Breeds

“…As part of a biodiversity conservation strategy, creating bioreservoirs of somatic and stem cell lines through cryogenic preservation can provide a reliable and feasible approach applied to regenerate or restore the biocenotic sustainability among herds of livestock breeds followed by reintroduction of vanishing breeds of farm animals into reconstituted niches of zootechnical biotopes [35,36]. Moreover, the establishment and subsequent cryoconservation of ex vivo-expanded permanent somatic or stem cell lines, which have been established from various tissue bioptates, serves as a promising tool independent of freezing or vitrification methods used for gametes and embryos [37][38][39].…”