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Research Article Volume 1 Issue 5
Culture, characterization and differentiation potential of rat bone marrow derived mesenchymal stem cells
Tamilmahan Paramasivam, Swapan Kumar Maiti, Sangeetha Palakara, Rashmi, Priya Singh, Naveen Kumar, Manjunthaachar, Christian Plank
Department of Veterinary Surgery and Radiology- Orathanadu, Tamilnadu Veterinary and Animal Sciences University, India
Division of Surgery, Indian Veterinary Research Institute, India
Division of Parasitology, Indian Veterinary Research Institute, India
Institute of Experimental Oncology and Therapeutics, Technical University of Munich, Germany
Received: July 30, 2016 | Published: October 17, 2016
Citation: Paramasivam T, Maiti SK, Palakara S, et al. Culture, characterization and differentiation potential of rat bone marrow derived mesenchymal stem cells. J Stem Cell Res Ther. 2016;1(5):193-199. DOI: 10.15406/jsrt.2016.01.00034
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 rBMC 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.
Keywords: rat bone marrow, mesenchymal stem cell, cd markers, osteogenic, adipogenic, chondrogenic, penicillin, alizarin red, alkaline phosphatase, alcian blue, distilled water, dried, calcium deposition, nanogram
©2016 Paramasivam, et al. This is an open access article distributed under the terms of the, which permits unrestricted use, distribution, and build upon your work non-commercially.