Experimental Study On PC12 Cells Induced And Differentiated Into Neurons

Object: To study the ability and the optimal concentration of pheochromocytoma cells(PC12 cells) differentiation into neurons induced by nerve growth factor(NGF), RA and BMSCs cultivated supernatant. PC12 cells would be constructed successfully for nervous protective function of NGF,RA and BMSCs cultivated supernatant.To make the PC12 cell line an extremely valuable neuronal model system.Method: Using morphological technique to observe the growth condition of PC12 cells which were exposed to different concentration of NGF, RA and BMSCs cultivated supernatant; Using immunohistochemistry technique to detect the expression of MAP2 in differentiated PC12 cells. Result: Undifferentiated PC12 cells are toroid,short fusiform and triangle,there are short neurites among some PC12 cells. Neurites have exserted after PC12 cells cultured for 24h,some of them are larger and bigger,similar to the neuronal axis.There are also some neurites inendochylema except neuronal axis,these neurites are different in shape and number. The length of total primary neurite and max diameter of cells in 50ng/ml NGF group and 1.0μg/ml RA group are obviously longer and bigger than that in control group when PC12 cells were cultured for 72h. There are some nets between PC12 cells.The higher of the concentration of NGF and RA, the larger of the number and lengh of the neurites when PC12 cells were exposed to 10,20 and 50ng/mlNGFand 0.1,0.3,0.5 and 1.0μg/ml RA.But unfortunately, the number and lengh of neurites are fewer and shorter when PC12 cells were exposed to 80ng/mlNGF and 2.0μg/ml or 5.0μg/ml RA than that of 50ng/ml NGF and 1.0μg/ml RA.There are a lot of black particles and cells became smaller and aging when PC12 cells were exposed 2.0μg/ml RA for 7d.The higher of the concentration of BMSCs cultivated supernament,the larger of the number and the lengh than that of control group. I also have used immunohistochemistry technique to detect the expression of MAP2 in differentiated PC12 cells except to observe the growth condition of PC12 cells which were exposed to different concentration of NGF, RA and BMSCs cultivated supernatant. The results show the number of MAP2 positive cells were significantly increased in 20, 50, 80ng/ml NGF and 0.3,0.5,1.0,2.0μg/mL RA and 40,80,120及160μl BMSCs cultivated supernament groups compared with control group. Especially the 50ng/mlNGF and 1.0μg/ml RA group. Conclusion: (1)PC12 cells could differentiate into neurons induced by NGF,RA and BMSCs cultivated supernament successfully. (2)50ng/ml is the optimal concentration in which NGF could induce PC12 cells to differentiate into neurons.(3)1.0μg/ml is the optimal concentration in which RA could induce PC12 cells to differentiate into neurons.