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.
Highlights
The ability of mesenchymal stem cells (MSCs) to differentiate into other mesoderm lineages such as bone and cartilage opened a variety of experimental strategies to investigate the possibility of these cells to be used in tissue engineering since MSCs derived from bone marrow (BM) have been applied in the treatment of musculoskeletal disease in several species [1]
The needle, anesthetic protocol, doses and MSCs processing was considerate appropriate for the species under study. From each animal it was collected on average 3 ml of BM and this volume was sufficient for the processing of the sample from these animals, promoting correct isolation, culture and characterization of the cells
The process of cooling the material during transport apparently did not affected the quality of the samples once all samples were submitted to culture
Summary
The ability of mesenchymal stem cells (MSCs) to differentiate into other mesoderm lineages such as bone and cartilage opened a variety of experimental strategies to investigate the possibility of these cells to be used in tissue engineering since MSCs derived from bone marrow (BM) have been applied in the treatment of musculoskeletal disease in several species [1]. Bone marrow-derived mesenchymal stem cells (BMMSCs) are the most studied source of MSCs and because of this, they have received special attention and are best characterized [2]. There are reports of studies with embryonic stem cells [9], MSCs derived from adipose tissue [1] and from amniotic fluid [10].
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