Comparison of Biological Features of Wild European Rabbit Mesenchymal Stem Cells Derived from Different Tissues

Abstract

Although the European rabbit is an “endangered” species and a notorious biological model, the analysis and comparative characterization of new tissue sources of rabbit mesenchymal stem cells (rMSCs) have not been well addressed. Here, we report for the first time the isolation and characterization of rMSCs derived from an animal belonging to a natural rabbit population within the native region of the species. New rMSC lines were isolated from different tissues: oral mucosa (rOM-MSC), dermal skin (rDS-MSC), subcutaneous adipose tissue (rSCA-MSC), ovarian adipose tissue (rOA-MSC), oviduct (rO-MSC), and mammary gland (rMG-MSC). The six rMSC lines showed plastic adhesion with fibroblast-like morphology and were all shown to be positive for CD44 and CD29 expression (characteristic markers of MSCs), and negative for CD34 or CD45 expression. In terms of pluripotency features, all rMSC lines expressed NANOG, OCT4, and SOX2. Furthermore, all rMSC lines cultured under osteogenic, chondrogenic, and adipogenic conditions showed differentiation capacity. In conclusion, this study describes the isolation and characterization of new rabbit cell lines from different tissue origins, with a clear mesenchymal pattern. We show that rMSC do not exhibit differences in terms of morphological features, expression of the cell surface, and intracellular markers of pluripotency and in vitro differentiation capacities, attributable to their tissue of origin.

Cryobanking European Mink (<i>Mustela lutreola</i>) Mesenchymal Stem Cells and Oocytes

“…Passaged cells had a more uniform morphology and developed into colonies as the culture developed. These morphological observations suggest that the isolated cells may contain both mature and progenitor populations, in line with what we have demonstrated in earlier studies on porcine, bovine, and rabbit species [29][30][31][32]. Our results show that all emMSCs isolated were positive for CD9, CD29, CD90, and CD44 characteristic mesenchymal markers not previously reported in European mink.…”

“…POU5F1, naturally expressed in embryonic and adult stem cells, has been identified as a key regulator of pluripotent stem cell differentiation and self-renewal [34]. Recent studies have reported the detection of POU5F1 or its transcription factor OCT4 in porcine [32], bovine [29,31], or wild European rabbit MSCs [30]. Although the MSC marker STRO1 has been reported in Ferret Dental Pulp Stem Cells, we were not able to detect its expression in none of the here characterized emMSCs.…”

“…Aside from cell biobanking, several studies in veterinary medicine have reported the improvement of chronic wounds by the action of combined treatments with MSCs [37] [38], representing a very interesting approach for possible treatments of animals in captivity. Oral mucosal MSC isolation has so far only been reported in humans [39] and rabbits [30]. We have reported here the isolation of five emOM-MSC.…”

“…Although the MSC marker STRO1 has been reported in Ferret Dental Pulp Stem Cells, we were not able to detect its expression in none of the here characterized emMSCs. SOX2, a pluripotency marker expressed in MSCs isolated from various tissues of different species such as red pandas [18,35], giant pandas [36], and wild European rabbits [30], was only observed in emO-MSC, OA-MSC, and emE-MSC. One of the main defining characteristics of a typical mesenchymal pattern is the ability of MSC to differentiate in vitro into mesodermal lineages [29][30][31][32], which was proved here for the European mink MSC isolated from different tissues.…”

“…For the isolation of primary cultures from the oral mucosa, dermal skin tissue, oviduct, endometrium, testis, bone marrow, and adipose tissue from different adipose depots: subcutaneous, ovarian, abdominal, and pericardial, the procedures have been detailed [29,30,32]. Briefly, the collected samples were minced before being incubated in 0.05% collagenase type II, 0.1% BSA, and 30 nM CaCl2.…”