Study of Chondrogenic Potential of Fibroblastic Cells Isolated From NMRI Mice

Purpose: The purpose of this study was to isolate and purify fibroblastic cells from NMRI mouse bone marrow and evaluate their potential in differentiating among condrocytic cell lineage in vitro. Materials and Methods: The cells from NMRI bone marrow were harvested and plated at about 500 cells per well of 6-well plates for several days. The pure fibroblastic cells were not appeared until the second passages of the cells were performed. To differentiate into cartilage, two different techniques were employed. In micro mass culture technique, about 200000 fibroblastic cells pletted by centrifuging were incubated in chondrogenic medium for three weeks. In Monolayer Culture technique, about 200000 cells seeded in 24-Well plate were provided with chodrogenic medium for three weeks. In the end of differentiation period, the cells were evaluated by histochemistry and RT-PCR analysis for cartilage differentiation. To examine the mesenchymal nature, the isolated cells were differentiated into bone and adipocytic cell lineages. Results: In the early days of primary culture, the dominant population of the cells was that with spindle shape morphology that expanded in subsequent subculture and almost purified in second passage.Differentiation results showed that in contrast to monolayer culture system in which chondrogenic differentiation was not occurred in micro mass system the cells were apparently differentiated into cartilage as it was evidenced by our evaluation. RT-PCR analysis was indicative of high production of collagen type II, X and aggreacan among the differentiated cells and histochemistry results showed the methachromatic matrix accumulation between the cells. The isolated cells in this study were mesenchymal cells as they readily differentiated into bone and adipocyte. Conclusion: The cells isolated by low-density culture system could be differentiated among the chondroblastic lineage and have mesenchymal characteristic.