[Experimental study on effect of monocyte chemoattractant protein 1 on migration of induced and differentiated mouse bone marrow mesenchymal stem cells in vitro].
OBJECTIVE
To investigate the effect of monocyte chemoattractant protein 1 (MCP-1) on the migration of the induced and differentiated mouse bone marrow mesenchymal stem cells (BMSCs) for raising the efficacy of intravenous transplantation of BMSCs.


METHODS
The BMSCs were cultured with the method of differential adhesion and density gradient centrifugation of C57/BL10 mice, and were identified by alkaline phosphatase Gomori modified staining after osteogenic inducing. At the 3rd passage, the BMSCs were induced to the myoblasts with 5-azacytidine (5-Aza). The chemotaxis of MCP-1 in the induced and differentiated BMSCs in vitro at concentrations of 25, 50, 100, 200, and 400 ng/mL was observed through the migration test, by counting the number of the migrated cells. The expression of the chemokine receptor 2 (CKR-2) in the induced and differentiated BMSCs was detected with the flow cytometry.


RESULTS
The cells could be cultured with the methods of differential adhesion and density gradient centrifugation and still had higher proliferative and differentiative potency; the induced cells at the 3rd passage could differentiate to the osteoblasts, confirming that the cells were BMSCs; the myogenic induced BMSCs possessed the sarcotubule structure. The number of the migrating BMSCs at MCP-1 concentrations of 25-400 ng/mL were respectively 35.066 7 +/- 6.584 2, 43.200 0 +/- 6.460 8, 44.466 7 +/- 4.823 5, 45.600 0 +/- 8.650 3, and 50.733 3 +/- 7.582 5; showing significant difference when compared with control group (28.333 3 +/- 8.917 6, P < 0.05), and presenting significant difference among 25, 50, 400 ng/mL groups compared with each other (P < 0.05). The expression of CKR-2 in the mouse BMSCs (48.0%) was significantly higher (P < 0.001) than those of blank control (0.6%) and negative control (17.0%).


CONCLUSIONS
The results indicate that the MCP-1 can induce the migration of mouse BMSCs by MCP-1/CKR-2 pathway.
Liu
Ju
J
Department of Neurology, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P R China.
Zhou
Hongyu
H
Liu
Libin
L
Luo
Li
L
Lian
Zhiyun
Z
Li
Huiying
H
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002465:Cell Movement; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000494:pharmacology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D032446:Myoblasts / Q000166:cytology
D018932:Chemokine CCL2