Lentiviral vector-mediated over-expression of CC chemokine receptor 7 promotes homing of rat bone marrow mesenchymal stem cells

doi:10.3969/j.issn.2095-4344.1789

Abstract

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells specifically express CC chemokine receptor 7 (CCR7). The interaction between secondary lymphoid-tissue chemokine and CCR7 contributes to the homing of bone marrow mesenchymal stem cells to the injured site.

OBJECTIVE: To explore the effects of CCR7 gene over-expression on the homing capacity of bone marrow mesenchymal stem cells in vivo.

METHODS: Rat bone marrow mesenchymal stem cells overexpressing CCR7 were constructed by lentiviral vector. We conducted a Transwell experiment to detect the effect of secondary lymphoid-tissue chemokine on the directional migration of bone marrow mesenchymal stem cells in vitro. Sprague-Dawley rats were divided into four groups (n=10/group): (1) Control group: non-irradiated rats were used as control group; (2) radiation group in which the rats were exposed to total body irradiation (⁶⁰Co, 0.75 Gy/min, 7.5 Gy in total), and then were infused with 1 mL of normal saline; (3) BMSCs-GFP group in which the rats were infused with bone marrow mesenchymal stem cell suspension (1 mL, 5×10⁵ cells) transfected by GFP via the tail vein after total body irradiation; (4) CCR7-BMSCs-GFP group in which the rats were infused with bone marrow mesenchymal stem cells (1 mL, 5×10⁵ cells) simultaneously carrying GFP and CCR7 gene via the tail vein after total body irradiation. The rats were sacrificed at 24 hours after infusion, and the frozen sections of the spleen were prepared to detect the distribution of infused cells. Furthermore, the homing rate of bone marrow mesenchymal stem cells to the rat spleen was detected by flow cytometry. The level of secondary lymphoid-tissue chemokine was detected by ELISA.

RESULTS AND CONCLUSION: (1) Results of the Transwell experiment showed that CCR7 over-expression gene promoted the directional migration of bone marrow mesenchymal stem cells to secondary lymphoid-tissue chemokine. (2) The CCR7 over-expression could significantly enhance the homing rate of bone marrow mesenchymal stem cells into the spleen. (3) Flow cytometry results slowed that the number of bone marrow mesenchymal stem cells overexpressing CCR7 homing into the spleen was significantly higher than that in the BMSCs-GFP group (P < 0.05). (4) ELISA results showed that the level of secondary lymphoid-tissue chemokine in the peripheral blood after 24 hours of irradiation significantly increased (P < 0.05). These findings indicate that the over-expression of CCR7 gene mediated by lentiviral vector can promote the homing of bone marrow mesenchymal stem cells into the spleen.

Key words: SLC/CCR7, bone marrow, mesenchymal stem cells, overexpression of CCR7 gene, stem cell transplantation, cell homing, spleen

CLC Number:

Wang Zhihong, Chen Weimin, Lin Yun, Shang Jin, Wei Tiannan.

Lentiviral vector-mediated over-expression of CC chemokine receptor 7 promotes homing of rat bone marrow mesenchymal stem cells

[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(25): 3973-3977.

Figures/Tables 1

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