施万细胞样细胞对大鼠背根神经节细胞突起生长和神经生长因子表达的影响及其机制

doi:10.13481/j.1671-587X.20220317

Abstract

Abstract: Objective

To investigate the effect of Schwann cell-like cells （SCLCs） on the growth of neurites and the expression of nerve growth factor （NGF） of dorsal root ganglion （DRG） cells in the rats， and to clarify the role of SCLCs on promoting the growth in the DRG cells and its mechanism.

Methods

The adipose-derived stem cells （ADSCs） were isolated and cultured，and the calcified nodules in the cells were detected by Alizarin red staining to observe their differentiation ability， and the ADSCs were induced to differentiate into the SCLCs.The DRG cells were isolated and cultured， and the expression of neuronal nuclear antigen （NeuN） protein in the DRG cells was detected by immunohistochemical staining. The co-culture system of SCLCs and DRG cells was established. The cells were divided into single culture group and co-culture group. HE staining was used to observe the morphology of the DRG cells in two groups， the number of DRG cells and the number of DRG cell processes in two groups were counted， immunohistochemical staining was used to detect the expression of NGF protein in the DRG cells in two groups， and Western blotting method was used to detect the expression level of NGF protein in the DRG cells in two groups.

Results

The primary ADSCs were cultured for 7 d， under the inverted microscope， a large number of cells with large volume and long spindle shape could be seen， which were similar to the arrangement of vortex or paving stone.The Alizarin red staining results showed that the calcified nodules can be seen under the microscope， and the cultured cells had the ability of differentiation. The immunofluorescence staining results showed that most cells were S100 staining positive， and the ADSCs could be successfully induced into the SCLCs. After cultured for 72 h， the processes of DRG cells were thin and long， arranged in a wreath-like arrangement. After 7－8 d， the number of cells was increased， the body of cells was large and round， and the processes were slender and connected like a fence. The immunohistochemical staining results showed that almost all the nuclei were marked as brown and NeuN staining positive. Compared with single culture group， the number of DRG cells and the number of DRG cell processes in co-culture group were increased significantly （P<0.05）， and the expression level of NGF protein in the DRG cells in co-culture group was increased significantly （P<0.05）.

Conclusion

SCLCs could promote the growth of DRG cells through increasing the number of DRG cells and neurites and upregulating the expression of NGF protein.

Key words: Schwann cell-like cells, Dorsal root ganglion cells, Co-cultivation, Nerve growth factor, Neuronal nuclear antigen

CLC Number:

R322.8

Yuanliang DU,Wang REN,Lin LIU,Shuodan HU,Mingyu LIU,Pengfei DU,Xiumei FU. Effect of Schwann cell-like cells on growth of neurites and expression of nerve growth factor in dorsal root ganglion cells of rats and their mechanisms[J].Journal of Jilin University(Medicine Edition), 2022, 48(3): 684-691.

Figures/Tables 6

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References 22

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Effect of Schwann cell-like cells on growth of neurites and expression of nerve growth factor in dorsal root ganglion cells of rats and their mechanisms

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