Study on differentiation of bone marrow mesenchymal stem cells into neurons induced by bone morphogenetic protein 7 in vitro
10.3760/cma.j.cn121113-20200227-00109
- VernacularTitle:骨形态发生蛋白7体外诱导大鼠骨髓间质干细胞向神经元分化的研究
- Author:
Guangchao BAI
1
;
Wen ZHANG
;
Lei GAO
;
Kuanxin LI
Author Information
1. 新疆生产建设兵团医院关节脊柱科，乌鲁木齐 830002
- From:
Chinese Journal of Orthopaedics
2020;40(12):802-810
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the role of BMP7 in inducing the differentiation of BMSCs into neurons in vitro.Methods:BMSCs were isolated and cultured by whole bone marrow adherence method. Adipogenic induction and osteogenic differentiation were used to test the multi-directional differentiation ability of BMSCs. BMSCs were randomly divided into control group and BMP7 groups (25 ng/ml, 50 ng/ml, 75 ng/ml and 100 ng/ml). The effect of BMP7 on the proliferation of rat BMSCs was measured by MTT assay. BMP7 induced morphological changes in rat BMSCS under an inverted phase contrast microscope. The relative expression levels of NF200, SYN1, MAP2 and GFAP mRNA in induced cells were measured by qRT-PCR. Immunofluorescence was used to measure the expression of NSE protein.Results:Rat BMSCs showed lipid droplets in the cytoplasm after adipogenic induction, and oil red O staining was positive; rat BMSCs showed opaque mineral nodules after osteogenic induction, and alizarin red stained was positive. At 2 d, 3 d, 4 d, 5 d, and 6 d after induction, the cell absorbance values of each group were statistically different ( P < 0.05). On the 6th day of induction, the absorbance values of the control group, 25 ng/ml BMP7 group, 50 ng/ml BMP7 group, 75 ng/ml BMP7 group and 100 ng/ml BMP7 group were 0.370±0.003, 0.399±0.003, 0.404±0.003, 0.410±0.003, 0.397±0.001, respectively. Cells absorbance value of 75 ng/ml BMP7 group was significantly higher than the other groups ( P< 0.05). The 75 ng/ml BMP7 group had the most significant changes in cell morphology, similar to neurons in cell morphology. The relative expression of NF200, SYN1, MAP2, and GFAP mRNA of 75 ng/ml BMP7 group (5.47±0.59, 1.48±0.38, 2.86±1.65, 4.41±0.13) was significantly higher than that of the control group ( P< 0.05). The positive rate of NSE immunofluorescence staining in the 75 ng/ml BMP7 group was higher than that in the control group (32.94%±1.62% vs 0). Conclusion:BMP7 has the ability to induce the differentiation of rat BMSCs into neurons in vitro.
