Bone marrow mesenchymal stem cells - protocol for isolation and cultivation from rats

Abstract

Mesenchymal stem cells (MSCs) can be obtained from different sources, such as adipose tissue, placenta, umbilical cord, etc., however, those obtained from the bone marrow are the main cell source used in tissue repair, tissue engineering and cell-based gene therapy. The methods of harvesting the bone marrow vary according to the specie. Usually, the smaller the subject, the more difficult is the harvesting and the number of cells obtained is more reduced. This paper describes a technique that we considered the most reliable and easy to apply for the isolation and differentiation of mesenchymal stem cells derived from rat bone marrow (BM-MSCs). For this purpose, humerus and femur bones from 16 weeks Wistar rat were used. The characterization of the cultured cells was performed by morphology evaluation using Olympus IX51 microscope and cell viability assay. The nucleated cells that attached to plastic exhibited a spindle shape morphology and all the passages showed a viability percent > 95, with a mean value of 96%. MSCs are emerging as a very promising therapeutic agent for tissue regeneration due to differentiation capacity and immunologic privilege. Their capacity to proliferate during numerous passages, making possible to obtain a large number is making MSCs very attractive from a therapeutic perspective.