The Conditioned Supernatant Of Mesenchymal Stem Cells On The Proliferation Effects Of PC12 Cell Model Of Parkinson Disease

Background Mesenchymal Stem Cells(MSCs) can play the role of neural repair and promote nerve regeneration by the mechanism of differentiating into neuron like cells, immune regulation and inducing nerve growth factor in situ to release [1-2] Granulocyte colony stimulating factor(G-CSF)can promote the neural differentiation angiogenesis and anti-inflammatory.It has obvious protective effect on the nervous system and also can promote the recovery of neural tissue’s structure and function. Parkinson’s disease(PD) is a kind of disease,which has a main pathological feature that the dopamin-ergic neurons of nigrostriatal pathway in midbrain have a degeneration. MSCs conditional medium was used by us to stimulate the proliferation of PD cell model of PC 12 cell, in order to verify nerve growth promoting effect of MSCs and speculate about the conditional medium containing G-CSF had stronger effect to promote the proliferation of experimen-tal cells.Objective Isolation, culture and identification of MSCs, to investigate the conditioned supernatanton of MSCs on the proliferation effects of PC 12 and it’s cell model of Parkinson disease.Methods We isolated long fusiform cells from umbilical cord tissue by tissue adherent method,identified the cells’immunophenotype by flow cytometry and their differentiation ability by staining method;treat the PC 12 cells with Mpp+to establish the PD cell model;the MSCs was stimulated by Concanavalin A(conA) and their conditioned supernatant was collected,then observed and compared the different effect of the conditio-ned supernatant and normal MSCs supernatant to the proliferation ability of PC 12 cell and it’s PD model and used ELISA to detect the content of granulocyte colony stimulating factor(G-CSF) in the two kinds of supernatant.Results1.Application of tissue adherent to separate the umbilical cord mesenchymal stem cells,giving the isolated cells a immune phenotype identification,which showed a high expression of stem cell surface markers CD 105, CD73, CD90 and did not express the hematopoietic stem cell surface markers CD 19, CD34 and CD45, this result accorded with the MSCs’ phenotype.2.1dentified the MSCs added Inducing agent conA,the phenotype of MSCs cells after induction did not change.3.Induced the isolated cells’adipogenic and osteogenic differentiation in different culture medium and gave them respectively ALP staining and Oil red O staining, the isolated cells In the experiment had good ability of adipogenic and osteogenic differentia-tion. The same method for identification showed the conA treated MSCs’osteogenic.. adipogenic differentiation ability had not changed obviously.4.The detection of CCK-8 kit showed that after the Mpp+ treatment,the number of PC 12 cells in the model group is half of the normal group.5.Compared with the control group,the MSC conditioned supernatant can improve the proliferation of PC12 and it’s cell model of PD,yet the MSC supernatant induced by conA could more significantly promote the proliferation, the difference had statistical signific-ance.6.After the detection of ELISA, the concentration of G-CSF in the MSCs supernatant induced by conA (33.52pg/mL) was significantly higher than that of the content of the ordinary MSCs supernatant(12.03pg/mL).Conclusion1.The isolated cells from experiment were mesenchymal stem cells,the induction of conA had not changed the MSC cytological characteristics.2.The PD cell model of PC 12 cells was successfully built.3.MSCs conditioned supernatant can promote the proliferation of PC 12 cell and it’s PD model,G-CSF may have significantly ability to promote the experimental cells’ proliferation.