Differentiation of Bovine Bone Marrow and Adipose Tissue Derived Mesenchymal Stem Cells Into Osteoblasts
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Date
2018-05-03
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Addis Ababa University
Abstract
Adipose tissue and bone marrow-derived stem cells have been extensively investigated for their applicability in the field of tissue engineering due to their multi-lineage differentiation potential. However, work on bovine-source adipose tissue and bone marrow-derived stem cells have been limited in use. In this work, bovine adipose tissue and bone marrow cells were cultured in osteogenic inductive medium for in vitro differentiation into osteoblast cells. Superficial white fat tissue and Bone marrow were isolated from freshly slaughtered 4,6-month-old cal. Cell proliferation as a measure of optical density was measured using absorbance reader at a wavelength of 570nm. Alizarin Red staining for the detection of calcium in the cultured cells was also performed. cDNA was synthesized from 20ng RNA. Using PCR, cDNA was amplified and its size was measured using Gel Electrophoresis. The morphologies of MSCs that proliferated in the in vitro culture were similar for cells from both adipose tissue and bone marrow. MTT assay test showed that the proliferation rate was lower for cells cultured in osteogenic medium compared to the cells cultured in the standard cell culture medium. All the gene markers for osteoblast cells were expressed by the MSCs cultured in the osteogenic medium. The culture induced osteoblast cells stained positive on Alizarin Red staining, indicating formation of calcified matrix. Differentiation of MSCs into osteoblast cells was better when combined with osteogenic medium. The study has provided evidence that adipose tissue MSCs can be used instead of embryonic stem cells and bone marrow MSCs as a source of osteoblast cell differentiation in vitro.
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Keywords
Mesenchymal Stem Cell, Bovine, Adipose Tissue, Bone Marrow, Osteogenesis