Differentiation of human umbilical cord-derived mesenchymal stem cells into annulus fibrosus-like cells by co-culture / 临床与实验病理学杂志

ABSTRACT

Purpose To induce the differentiation of hu-man umbilical cord mesenchymal stem cells ( HUCMSCs) into annulus fibrosus (AF) cells by in vitro co-culture technique and to investigate the morphological and histological changes of HUCMSCs after co-culture. Methods HUCMSCs and AF cells were isolated from the normal neonatal umbilical cord and New Zealand white rabbit. Transwell six-well plates were used for co-culture with the cells seeded at the ratio of 1 ∶ 1, HUCMSCs cultured alone served as controls. After two weeks of co-culture, morphological changes were observed by inverted microscope. Real-time PCR was used to detect the expression of typeⅠcolla-gen, aggrecan and SOX-9 gene in HUCMSCs. Immunocyto-chemical staining and toluidine blue staining were used to detect the synthesis of cell matrix such as type Ⅰ collagen and aggre- can. Results The morphology of HUCMSCs in control group was long-fusiform, the morphology of HUCMSCs in co-culture gradually became short-fusiform or polygonal, and began to ap-pear synapse, showing the morphological features of AF-like cells. Real-time PCR results showed that typeⅠcollagen, aggre-can and SOX-9 mRNA were significantly increased in the co-cul-ture group (P＜0. 05). Immunocytochemical staining and tolui-dine blue staining showed that type I collagen and aggrecan were positive, respectively. Conclusion In vitro co-culture technol-ogy can induce HUCMSCs to differentiate into AF-like cells, which is expected to provide a new kind of seed cells for the bio-logical treatment of degenerative disc disease.