动物造模,RSC96 z-bio 2021-01-29 310

　　Objective: To observe the effect of conditioned medium on rat Schwann cell line RSC96 cells (RSC96-CM) on the proliferation of oligodendrocyte progenitor cells (OPC) and to explore its mechanism of action.

　　Method: Isolate OPC from the embryonic spinal cord of SD rats by immunoadhesion method within 15 days, detect the proliferation of OPC by BrdU uptake experiment, and perform RT-PCR in RSC96 cells to detect the expression of PDGF-AA and bFGF. Use ELISA technology to detect the levels of PDGF-AA and bFGF protein in RSC96-CM, observe the effect of PDGF-AA and bFGF on the growth of OPC induced by specific inhibitors of RSC96-CM, and observe ERK and JNK signals to observe the influence of this pathway . The role of SC96-CM in inducing the growth of certain inhibitors of OPC.

　　Result: Compared with the control group, the proportion of BrdU + cells of OPC cultured with different concentrations of RSC96-CM increased significantly (P\u003c0.05). Among them, OPC cultured in a medium containing 50% RSC96-CM reaches a peak when the proportion of BrdU + cells increases significantly. T-PCR confirmed that RSC96 cells significantly expressed PDGF-AA and bFGF mRNA. According to the ELISA results, it was found that the protein concentration of PDGF-AA and bFGF of RSC96-CM was 0.87, which is the protein level of PDGF-AA and bFGF of B104CM. Previous report. Times and 0.92 times. Both the specific inhibitor of PDGFRAG1295 and the specific inhibitor of bFGFRPD173074 significantly reduced the OPC proliferation induced by RSC96-CM. In addition, Erk1/2 specific inhibitors U0126 and JNK specifically inhibit the pre-incubation of SP600125, and also significantly reduce the BrdU + OPC ratio induced by RSC96 and -CM (P\u003c0.01).

　　 Conclusion: RSC96-CM significantly induces OPC proliferation and activates Erk 1/2 and JNK signaling pathways through PDGF-AA and bFGF secreted by RSC96. SC96-CM can also be used as an amplifying agent in conventional cultures.