Autologous Platelet Lysate Is an Alternative to Fetal Bovine Serum for Canine Adipose-Derived Mesenchymal Stem Cell Culture and Differentiation

SIMPLE SUMMARY: Fetal bovine serum is extracted from cow fetuses and is the primary source of nutrition for cell growth in laboratory cell culture media. Exclusively for research purposes, the fetal bovine serum is harmless, but for transplantation research, it can cause severe immune disease in the host. Keeping this serious ethical and scientific issue in mind, we used allogeneic platelet lysate, a nutritional supplement obtained from the same animal that will be transplanted with its stem cells, for stem cell culture. Cell culture media supplemented with different combinations of fetal bovine serum or allogeneic platelet lysate were used in this study. Parameters such as cell doubling time, cell viability, stem cell expression markers, and gene expression of various markers related to the transformation of stem cells into adipocytes and bone cells. Our results showed that with the use of an autologous platelet lysate, the values of all parameters were far better when compared to fetal bovine serum. This means that autologous platelet lysate can be used as a substitute for fetal bovine serum without changing the differentiation potential of cells, thereby addressing the ethical and scientific issues of fetal bovine serum for regenerative medicine. ABSTRACT: The use of fetal bovine serum (FBS) in regenerative medicine raises serious ethical and scientific concerns. We have cultured and differentiated the canine mesenchymal stem cells (cMSCs) in five different media combinations of autologous platelet lysate (A-PL) and FBS; consisting of 0% A-PL and 10% FBS (M-1), 2.5% A-PL and 7.5% FBS (M-2), 5% A-PL and 5% FBS (M-3), 7.5% A-PL and 2.5% FBS (M-4), and 10% A-PL and 0% FBS (M-5). The cMSCs were evaluated for their doubling time, differentiation efficiency, and expression of CD73, CD90, CD105, and PDGFRα. The mRNA expression of NT5E, THY1, ENG, PPARγ, FABP4, FAS, SP7, BGLAP, and SPP1 was also assessed. The results indicated non-significant differences in cellular proliferation/viability; positive expression of surface markers, and PDGFRα with substantial adipo/osteogenic differentiation. The expression of adipogenic (PPARγ, FABP4, FAS), and osteogenic (SP7, BGLAP, SPP1) genes were higher (p < 0.05) in the M5 group. In conclusion, A-PL in cMSCs culture did not negatively affect cellular proliferation and viability but also enhanced their genetic potential for multilineage differentiation. Our results indicate that A-PL can be used as an alternative for FBS to develop potent cMSCs under good manufacturing practice protocol for regenerative medicine.