Homing of Cultured Endothelial Progenitor Cells and Their Effect on Traumatic Brain Injury in Rat Model

Xin-Bin Guo,Ying Wei,Xin Deng

doi:10.1038/s41598-017-04153-2

Xin-Bin Guo, Ying Wei + Show 1 more

Open Access

https://doi.org/10.1038/s41598-017-04153-2

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Abstract

Transplanted endothelial progenitor cells (EPCs) may play an important role in reestablishing the endothelial integrity of the vessels after brain injury, and contribute to neurogenesis. We, therefore, tested the homing of ex vivo cultured peripheral blood-derived EPCs and their effect on injured brain tissue after intravenous administration. To track the homing of implanted EPCs in injured brain tissues, EPCs were labeled with DAPI and BrdU in vitro before transplantation. EPCs were transplanted into the host animal through peripheral administration through the femoral vein, and homing of EPCs was evaluated. The integration of intravenously injected EPCs into the injured brain tissue was demonstrated. Immunohistochemical staining showed that microvessel density in the perifocal region of EPCs-transplanted rats was significantly increased, and the numbers of BrdU+ cells in the DG of subventricular zone were increased in EPCs-transplanted rats as compared to the control group. Transplanted EPCs may play an important role in reestablishing the endothelial integrity in the vessels after brain injury and further contribute to neurogenesis. EPCs enhanced recovery following brain injury in a rat model of TBI.

Highlights

The exact roles of transplanted endothelial progenitor cells (EPCs) in reestablishing the endothelial integrity of vessels after brain injury and their therapeutic values in neural recovery remain unclear
The cells were suspended in EBM-2 medium (Clonetics, USA) supplemented with 10% fetal calf serum (FCS, Xiamen Tebao Bioengineering Company), vascular endothelial growth factor (VEGF), human fibroblast growth factor-B, human epidermal growth factor, and plated on rat-derived 10 μg/ml fibronectin-coated plates (Sigma Chemical)
After the induction of TBI, rats were injected with DAPI and BrdU labeled EPCs (1 × 106 cells/rat) to test the homing and unlabeled EPCs to assess their effect on the neurogenesis, angiogenesis and the neurobehavior by tail vein injection

Summary

Materials and Methods

All experimental procedures were approved by the Care of Experimental Animals Committee of Zhengzhou University and performed in accordance with the relevant guidelines and regulations. To test the homing of transplanted and labeled EPCs, TBIs were induced using a fluid percussion injury device (MODEL01-B, NEW SUN, CA, USA) in eight rats. The EPC-transplanted rats were administered BrdU (75 mg/kg every 6 hour)) intraperitoneal[1] day after the transplantation of unlabeled EPCs,and PBS was injected into the peritoneal cavity of the control group[12]. Transplantation of ex vivo expanded EPCs. After the induction of TBI, rats were injected with DAPI and BrdU labeled EPCs (1 × 106 cells/rat) to test the homing and unlabeled EPCs to assess their effect on the neurogenesis, angiogenesis and the neurobehavior by tail vein injection. The mNSS test was performed in the control and EPCs-treated groups (12 rats in each group) prior to and on the 1st, 3rd, 5th, 7th, 14th, 21st, and 25th days after the TBI procedure.

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