Description of the technique of bone marrow harvesting in the coxal tuberosity for isolation and culture of mesenchymal stem cells of buffaloes (Bubalus bubalis)

Abstract

Several studies with mesenchymal stem cells (MSCs) have been developed in many species because of its ability to differentiate into other mesoderm lineages, capacity of self-regeneration, low immunogenicity, paracrine, anti-inflammatory, immunomodulatory and antiapoptotic effects which make them a promissory source to be used in therapeutic strategies. The aim of this study is to report the technique of harvest of bone marrow (BM) in the coxal tuberosity (CT) of buffaloes and its processing and cultivation. For this, after anesthetic block from the region corresponding to the CT, bone marrow harvesting was performed with a myelogram’s needle. The samples collected showed plastic adherence with 96 hours and took approximately 32 days to reach 80% confluence. And then differentiation into adipogenic and osteogenic lineages was performed. Samples showed morphological changes during differentiation protocol, but not all presented production of extracellular deposits of calcium or intracellular fat droplets. The anatomical site tested showed to be an alternative site of harvest of BM once provided with the appropriate isolation and culture of the mononuclear fraction. Moreover, the procedure was performed without difficulty and with great security. Based on this, it can be concluded that CT is an excellent anatomical site for isolation and culture of MSCs and the proposed technique is viable and feasible to be held in buffaloes.