Adipose Tissue- and Bone Marrow-Derived Mesenchymal Stem Cells from Sheep: Culture Characteristics

Abstract

The current study demonstrates the culture characteristics of adipose tissue and bone marrow-derived mesenchymal stem cells (MSC). The study evaluates the effect of ambient temperature, physiological status of the donor and the tissue source on sheep (Ovis aries) mesenchymal stem cells. The tissue samples were harvested from full term pregnant female sheep (n = 9) and male sheep (n = 10). Adipose tissue was harvested from n = 9 sheep and bone marrow from n = 10 sheep. The samples (adipose tissue, n = 2; bone marrow, n = 3) transported at cold ambient temperature (<10 °C) failed to yield MSCs while those (n = 14) at higher (>20 °C) ambient temperature successfully yielded MSCs. Bone marrow mononuclear cell (MNC) fraction was higher than the adipose tissue-derived stromal vascular fraction (SVF), but the percent adherent cells (PAC) was higher in the later cell fraction. Adipose tissue-derived MSCs from the full term female sheep had a significantly (p < 0.05) higher proliferation potential as compared to those of the male sheep-derived MSCs. Female sheep MSCs also had rapid differentiation potential. The cryopreserved MSCs had morphological features comparable to that of the fresh cells. In conclusion, the tissue type and physiological status of donor animal may affect MSCs’ characteristics and should be taken into consideration while applying in clinical settings.

Clinical applications of adipose-derived stromal vascular fraction in veterinary practice

“… 2021 ; Dar et al. 2021 ). The mononuclear cell (MNC) fraction harvested contains MSCs that are culture expanded.…”

“…2016 ; Dar et al. 2021 ). One of the studies demonstrated that AdSVF composes mainly of AdMSCs (15–30%) along with other cellular components like immune cells (25–45%), endothelial cells (10–20%), and pericytes (3–5%) (Bourin et al., 2013).…”

“…Hence, the number of AdMSCs present in adipose tissue will range from 5000 to 200,000 stem cells/g (Baer and Geiger 2012 ; Dar et al. 2021 ). The composition of the AdSVF, as well as the proliferation rate and differentiation capacity of the AdSVF cells, depends on factors such as animal species, age, type (brown or white) and anatomical location (subcutaneous or visceral) of adipose tissue, type of surgical procedure, method of cell separation, culturing conditions, culture medium, exposure to plastic, and plating density (Gentile et al.…”

Multi-faceted characterization of mesenchymal stem cells and their application as a powerful tool for agrobiotechnology, assisted reproductive technologies, and veterinary and regenerative biomedicine – a review

“…Although the immunophenotypic landscapes of ovine MSCs are similar to human MSCs, most antibodies reacting with human MSCs did not exhibit the reactivity with the surface antigens of ovine MSCs. The investigations performed by Dar et al (2021) have shown that ovine BM-and ATderived MSCs are simultaneously characterized by the increased expression levels recognized for CD73 and CD90 markers, and the absence of CD34 and CD45 antigens. Other studies undertaken to identify the immunophenotypic landscapes specific for ovine BM-MSCs have confirmed the high expression of CD29, CD166 and CD44 antigens and the lack of expression of the CD45 marker (Sanjurjo-Rodríguez et al, 2017).…”

Connection between Mesenchymal Stem Cells Therapy and Osteoclasts in Osteoarthritis

“…[134]. Indeed, many factors, such as the storage temperature of the MSCs and preservation agents employed, could also affect the effectiveness of this cell therapy [135,136]. For these reasons, we are still far from knowing the most optimal conditions for the use of MSCs as a therapeutic tool in preclinical and clinical trials in OA [137,138].…”