动物造模 z-bio 2022-07-19 374

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

　　METHODS: OPCs were isolated from the spinal cord of 15d embryonic SD rats by immunoadhesion method, the proliferation of OPCs was detected by BrdU incorporation assay, the expression of PDGF-AA and bFGF in RSC96 cells was detected by RT-PCR, and RSC96 was detected by ELISA. -The concentration of PDGF-AA and bFGF protein in CM, the role of PDGF-AA and bFGF in RSC96-CM-induced OPCs proliferation was observed with specific inhibitors, and the ERK and JNK signaling pathways were observed with specific inhibitors in RSC96- The role of CM in the induction of OPCs proliferation.

　　Results: Compared with the control group, the proportion of BrdU+ cells of OPCs cultured with different concentrations of RSC96-CM was significantly increased (P<0.05). Among them, the BrdU+ cells of OPCs cultured in the medium containing 50% RSC96-CM ratio peaked. RT-PCR confirmed that RSC96 cells significantly expressed PDGF-AA and bFGF mRNAs, and ELISA results showed that the protein concentrations of PDGF-AA and bFGF in RSC96-CM were 0.87 of the protein levels of PDGF-AA and bFGF in B104CM that we reported earlier, respectively. times and 0.92 times. Both the PDGFR-specific inhibitor AG1295 and the bFGFR-specific inhibitor PD173074 significantly reduced RSC96-CM-induced OPCs proliferation; in addition, Erk1/2-specific inhibitor U0126 and JNK-specific inhibitor SP600125 preincubation also significantly reduced RSC96-CM Proportion of induced BrdU+ OPCs (P<0.01).

　　CONCLUSION: RSC96-CM significantly induces the proliferation of OPCs, which plays a role through the activation of Erk1/2 and JNK signaling pathways by PDGF-AA and bFGF secreted by RSC96; RSC96-CM can also be used as an expansion agent for conventional culture of OPCs.