Survival of the proliferation bone marrow mesenchymal stem cells transplaned into rats infarcted rat brain
10.3969/j.issn.1672-5921.2009.09.007
- Author:
Yang WANG
1
Author Information
1. Department of Neurology
- Publication Type:Journal Article
- Keywords:
Ascorbic acid;
Cell proliferation;
Fibroblast growth factor 2;
Mesenchymal stem cells;
Rats;
Transplantation
- From:
Chinese Journal of Cerebrovascular Diseases
2009;6(9):475-480
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To study the effects of vitamin C and basic fibroblast growth factor (bFGF) on proliferation in vitro and transplantation in vivo of bone marrow mesenchymal stem cells (BMSCs) in rats with cerebral infarction. Methods: Circled digit oneBMSCs of rats were isolated by using density gradient centrifugation and adherent method. The rats were divided into four groups according to the different growth factors added in the medium: control group, Vc group (50 μg/mL), bFGF group (1 μg/L), and Vc + bFGF group (50 μg/mL Vc and 1 μg/L bFGF). The cellular morphology in each group was observed. MTT colorimetric method was used to detect the absorbance (A value) at the wavelength of 450 nm at day 1, 2, 3, 5, and 7 after the culture; flow cytometer was used to detect the cell cycle cultured after 96 hours. Circled digit twoA model of right middle cerebral artery occlusion (MCAO) for 2 hours was induced by the suture method in 20 rats, and they were randomly divided into 2 groups: the BMSC transplantation group cultured by the combined Vc + bFGF and the control BMSC transplantation group. BMSCs in the corresponding groups were transplanted in rats 24 hours after cerebral infarction. At 1, 2, and 3 weeks after transplantation, the brain tissue slices of the rats were prepared, and they were revealed by BrdU immunohistochemistry staining. Results: Circled digit oneMorphological observation showed that the proliferation of BMSCs was most fast in the Vc + bFGF group, and it was relatively slow in the control group. Circled digit twoThe A value of cells in each group began to increase at day 2 after the culture. It reached the peak at day 3 in the Vc group and bFGF groups (F = 728.52 and F = 197.18, P < 0.05); it reached the peak at day 5 in the Vc + bFGF group (F = 1771.32, P < 0.05), and then both deceased at day 7. From the second day, the A value in the bFG, Vc and Vc + bFGF groups was significantly higher than that in the control group; from the third day, Vc + bFGF group was higher than the Vc and bFGF group, and there was significant difference (P < 0.05). Circled digit threeThe percentage of cells in the proliferative phase (S + G2 + M) in the Vc, bFG and Vc + bFGF groups was higher than that in the control group, and there was significant difference (P < 0.05). Circled digit fourTwo weeks after the BMSC transplantation, BrdU-positive cell counts in the brain tissue slices in the BMSC transplantation group cultured by the combined Vc + bFGF was higher than that in the control BMSC transplantation group; the positive cell counts in both groups decreased at week 3, however, the former was still higher than the latter, and there was significant difference (P < 0.01). Most BrdU labeled BMSCs appeared in the centre or around the infarct brain. Conclusion: Both Vc and bFGF may promote proliferation of BMSCs in rats. The efficacy of the combination of both is better than any one alone. Survival ability of the BMSCs are obviously strengthened when they are transplanted into rats infarcted brain.
