Characterization of Rabbit Mesenchymal Stem/Stromal Cells after Cryopreservation

SIMPLE SUMMARY: Mesenchymal stem cells (MSCs) are prime candidates for cell-based therapies and regenerative medicine due to their trilineage differentiation potential. MSCs can differentiate into osteocytes, adipocytes, and chondrocytes. A significant number of these cells must be available for clinical application. As a result, cryopreservation becomes essential to ensure the cells are readily available and can be stored long term. However, there is a gap in our knowledge concerning the characteristics, sustainability of multipotency, and biosafety of cryopreserved MSCs in rabbits. Additionally, the functional ability of MSCs varies based on their source, which could influence their cryopreservation response. Differences in morphology, differentiation potential, and resilience to cryopreservation might also exist among various animal models and humans. To learn more, we assessed MSCs’ characteristics and functional properties from rabbits before and after cryopreservation. These cells were sourced from adipose tissues and bone marrow. Our findings suggest that while bone marrow-derived MSCs retain their functional ability poorly, adipose-derived cells retain their functionality better. ABSTRACT: Adipose tissues (ADPs) are an alternative source for mesenchymal stem/stromal cells (MSCs), given that conventional bone marrow (BM) collection is painful and yields limited cell numbers. As the need for easily accessible MSCs grows, cryopreservation’s role in regenerative medicine is becoming increasingly vital. However, limited research exists on the characteristics and functional properties of rabbit-derived MSCs from various anatomical sources before and after cryopreservation. We examined the effects of cryopreservation using Bambanker. We found that cryopreservation did not adversely affect the morphology, viability, and adipogenic or chondrogenic differentiation abilities of ADP MSCs or BM MSCs. However, there was a notable drop in the proliferation rate and osteogenic differentiation capability of BM MSCs post-cryopreservation. Additionally, after cryopreservation, the surface marker gene expression of CD90 was not evident in ADP MSCs. As for markers, ADIPOQ can serve as an adipogenic marker for ADP MSCs. ACAN and CNMD can act as chondrogenic markers, but these two markers are not as effective post-cryopreservation on ADP MSCs, and osteogenic markers could not be validated. The study highlights that compared to BM MSCs, ADP MSCs retained a higher viability, proliferation rate, and differentiation potential after cryopreservation. As such, in clinical MSC use, we must consider changes in post-cryopreservation cell functions.