Bone marrow mesenchymal stem cells induced by glial cell line-derived neurotrophic factors differentiate into neuron-like cells in vitro

doi:10.3969/j.issn.2095-4344.1818

Abstract

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells are pluripotent stem cells in mesoderm, not only differentiating into mesenchymal lineage cells such as osteoblasts, adipocytes and chondrocytes, but also having a neural differentiation potential. Therefore, they have a wide range of applications.
OBJECTIVE: To investigate the feasibility of bone marrow mesenchymal stem cells differentiating into neuron-like cells after in vitro induction with glial cell line-derived neurotrophic factor.
METHODS: Bone marrow mesenchymal stem cells from healthy Sprague-Dawley rats were cultured and purified in vitro using the whole bone marrow culture method. Cell phenotypes were identified using flow cytometry. Passage 4 bone marrow mesenchymal stem cells growing well were assigned into control group with no induction or experimental groups with addition of 20, 50, and 100 μg/L glial cell line-derived neurotrophic factors. Cell growth and morphology were continuously observed by inverted phase contrast microscope. Expressions of nestin and neuron-specific enolase were identified by immunocytochemical technique at 6, 12, 24, 48, and 72 hours of induction.
RESULTS AND CONCLUSION: (1) Primary cultured bone marrow mesenchymal stem cells appeared with high morphological uniformity and strongly expressed CD44 and CD90, and lowly expressed CD34 and CD45. (2) After induction with glial cell line-derived neurotrophic factors, the cell body gradually shrank towards the nucleus, accompanied by cell deformation that the cells presented with the typical morphologies of neuron-like cells, such as dipolar, multipolar and pyramidal. Cell protuberances extended and were gradually interconnected with adjacent cells. (3) The cells were positive for nestin at 6 hours of induction with glial cell line-derived neurotrophic factors, and the positive expression peaked at 24 hours of induction. Neuron-specific enolase expressed positively at 12 hours of induction and the expression peak appeared at 48 hours of induction. The expression levels of nestin and neuron-specific enolase were significantly higher in the 50 and 100 μg/L glial cell line-derived neurotrophic factor group than the 20 μg/L glial cell line-derived neurotrophic factor group. There were no obvious morphological changes and specific marker expressions of neuron-like cells in the control group. To conclude, rat bone marrow mesenchymal stem cells isolated by the whole bone marrow culture method can be cultured primarily and subcultured in vitro, and have the potential to differentiate into neuron-like cells induced by glial cell line-derived neurotrophic factor.

Key words: bone marrow mesenchymal stem cells, glial cell line-derived neurotrophic factor, cell differentiation, nerve cells, nestin, neuron-specific enolase, National Natural Science Foundation of China

CLC Number:

Liu Zhendong, Wang Rui, Yang Jiandong, Wang Jingcheng. Bone marrow mesenchymal stem cells induced by glial cell line-derived neurotrophic factors differentiate into neuron-like cells in vitro[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(29): 4605-4609.

Figures/Tables 1

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