[Differentiation of somatic unrestricted cord blood stem cells into chondrocyte cells]

ABSTRACT

To isolate and purify unrestricted somatic stem cells from human umbilical cord blood and evaluation of their differentiation into chondrocyte in vitro. In this study cells from human umbilical cord blood were isolated and plated in flask. Colonies were performed after one week. To determine the kind of cells, 100000 cells were analysed with flowcytometry. Twenty thousands [200000] of cells were plated in 6 wells that coated with poly-L-lysine II and incubated in chondrogenic medium to analyze the differentiation of these cells into cartilage. After 24 hs the first pletted cells were formed that continued to be existing to 21 days. The culture of cells were exchanged into chondrogenic culture every two days. At the end of differentiation period and 3 weeks the cells were analyzed by Alcian blue, immunohistichemistry and RT-PCR In addition these cells were passaged 50 times and their karyotyping analyzed. In early days of primary cultures, the number of spindle cells were increased and almost purified in second passage. The differentiation by RT-PCR analysis showed high production of collagen II, aggrican, BMP-6 and collagen that all are the specific genes of chondrocye cells, and.histochemistry assay showed that the methachromatic matrix was accumulated between the cells and expression of collagenll was confirmed. Karyotyping analysis showed high passages for these cells that was expected. Cultured USSC Differentiated into a chondroblast cell linage potential source for cell transplantation for rheumatoid arthirits as soon as cord blood is better source for mesenchymal Stem cells against bone marrow