| BackgroundRepair of articular cartilage defect is always a very difficult problem in the field of surgery. In past ,most studies used periosteum cell,cartilage or direct repair of cell transplanting, but it couldn't lead to cell division, proliferation , mesenchymal synthesization and cartilage forming , so that it used to fail .The itself cell source is limited and easy to age and de-differentiation when prolifen plantly in vitro, it is difficult develop cartilage tissue, and absent mechanics and tolerance of normal clarity cartilage. While self-cartilage cells transplantation.Repairs the default tissue, another default tissue forms after transplantation. Man-made tissue substitute is easy to induce repulsion reaction. Sponsoral infection, moved out at last. Induery Mscs into chondrocytes is regarded as practical method to therapy articular cartilage defect by tissue engineering.We can get different quantity and quality mesenchymal cells because different inducing methods, on tusside, their expressive way of gene and protein expression are different, we can exam their expressive difference by gene chip, we can test notable different membrane marker of gene expression. special adhere, by Flow Cytometry. By analysis the study show what factors take part in during differentiation. Objective:Based on the result of different revrulsants or revulsant combination which induced MSCs into chondrocyte obtained by the methods of morphology, imunohisto chemistry, in situ hybridization and RT-PCR, we detected gene expression profile of MSCs adhere factors in the process of different tiating into chondrocy at vary situation including different revulsant or revulsant combination. Regarded MSCs cultured without any revulsant as control group, wecanalized and compared the difference of vary group expression; we used to detect the MSCs membrane marker based on different gene profile. To discuss the mechanism n best cultural condition and protein expression of MSCs induced into pre-chondrocyte .We expected to find out the mechanism of correlative gene expression during MSCs into chondrocyte.In this study, we designed three contents as follows : ( 1 ) To detect the marker of chondrocyte by morphology ? in situ hybridization and RT-PCR in order to identify if MSCs differentiate to chondrocyte, at the same time, to validate the effect of different revulsants or revulsant combination. (2) To detect gene expression profile of MSCs adhere by gene chip in the process of differentiating into chondrocyte at the situation of different revulsants or revulsant combination. ( 3 ) To detect the marker of chondrocyte induced by different revuJsants or revulsant combination by Flow Cytometry in order to identify if gene expression of cell adhesive receptors accordant to marker of chondrocyte. Result1. Morphologic observation and marker of MSCs into chondrocyte in vitroUsing morphology to observe cell configuration ,to detect collagen II expression by RT-PCR. Induced by DEX and TGF: P ,morphology of MSCs not only had topical character of cartilage cells but also had the most bloom mesenchymal sersted grains, expression of collagen II was strongest and their proliferation was best.2. To analyze gene expression difference of MSCs to chondrocyte MARs by gene chipIn 2252 target genes, we did not find out gene diversity expression of MARs protein in group A; In group B,X06564,Y13714,X16563,and M31725 were down-regulated, without up-regulation; In group C, there seven diversity genes, including five up-regulated and two down-regulated, the expression of M31725 was consistent with expression of group-$ ;There were four diversity genes up-regulated in group D.3 .The result of Flow CytometryInduced by DEX and TGF- P j,CD56 expression of MARs was higher than other groups. ConclusionBy gene expression profile and Flow Cytometry, gene expression of MARs protein up-regulated and differentiated mature marker in the group which induced by DEX and differentiated mature marker and TGF- Pi. It showed that high expression of MARs protein and differentiated mature marker perhaps took part in the process of MSCs to chondrocyte. The result was consistent with morphology *. in situ hybridization and RT-PCR. It was more in favor of differentiating into chondrocyte using TGF-B and proper dosage DEX to treat MSCs than TGF- ï¿¡ i alone. |
