Culture, characterization and differentiation potential of rat bone marrow derived mesenchymal stem cells

Abstract

Bone marrow derived stem cells could offer significant multipotent differentiation capacity and plasticity in other species. However, in rat marrow there are few reports. This study described, simple and easy method for isolation of rat bone marrow stem cells (rBMSC) based on their density and plastic adherence properties and along with their trilineage differentiation potential. Rat bone marrow stem cells were isolated by density gradient and plastic adherence method. After isolation of stem cell they were characterized by phase contrast microscopy and flow cytometry. Moreover, differentiation into osteoblast, adipocytes and chondrocytes of mesenchymal stem cells (MSC) on different induction medium was analyzed using RT-PCR and their respective staining methodology. The attached nucleated cells showed their spindle shaped fibroblastic like morphology. Flow cytometry results revealed that they were negative for haematopoietic markers such as CD 31 and CD 45 and positive for mesenchymal markers CD 90 and CD 29. On the other hand, they were differentiated into osteogenic, adipogenic and chondrogenic and this was confirmed by alizarin red, Oil red O and Alcian blue stain respectively. Further RT-PCR results indicated that rBMSC expressed Runx2; Ppar-γ and Aggrecan genes which showed that these cells can easily differentiate into many different lineages and this can be applicable for cell based regenerative disorders.