Murine mesenchymal stem cell isolation: impact of cell-seeding density on morphology, differentiation and profile of surface antigens
Trauma Monthly: 13 (1); 37-49 Article Type: Research Article
S. Murine mesenchymal stem cell isolation: impact of cell-seeding density on morphology, differentiation and profile of surface antigens,
Online ahead of Print
Objective: The aim of the present study was to isolate MSCs using either low or high cell-seeding density culture system and to compare the cells produced in two systems in terms of their morphology, differentiation potential as well as the expression of some cell surface antigens. Material and Methods: The bone marrow cells from Balb/c mice of 6-8-weeks old were cultivated at 2.5×106 cells/cm2. Primary culture was then tripsinized, cultured either at 50 cells/cm2 as low density culture systems or at 8×104 cell/cm2 as high density culture system. The passaged-3 cells from either system were further examined and compared in terms of morphology, differentiation potential and the expression of some surface antigens including CD135, CD44, CD31, Thy1.2, CD11b, CD45, CD34, Vcam1, Sca-1, and c-Kit. Results: In contrast to high–density culture system, in low density one, a few colons were produced and the culture contained more fibroblastic cells. According to the differentiation results, in contrast to high density system, more percentage of the cells being produced in low density culture system has been differentiated into bone and adipose cells. Furthermore, CD135, CD34, CD31 and Vcam were not appeared on the cells of low density system and Thy 1.2 surface antigen was not expressed on the cells of high density culture system. All these differences were statistically significant. Conclusions: Taken together, it seems that low density culture system is an appropriate method to isolate and expand murine mesenchymal stem cells.
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