Fluorescence tracing of umbilical cord blood mesenchymal stem cells in transplantation treatment of scalded wound in mice
10.3969/j.issn.1673-4130.2019.04.011
- VernacularTitle:脐血间充质干细胞经转染绿色荧光蛋白后治疗SCID小鼠烫伤的创面效应示踪
- Author:
Liyuan QU
1
;
Mengwen LI
;
Zhenming LI
;
Xigui PAN
;
Xin WANG
Author Information
1. 解放军第970医院/原解放军第107医院输血科
- Keywords:
mesenchymal stem cell;
transplantation;
enhanced green fluorescent protein (GFP);
fluorescence tracing
- From:
International Journal of Laboratory Medicine
2019;40(4):423-426
- CountryChina
- Language:Chinese
-
Abstract:
Objective Using the previously established mesenchymal stem cells strain derived from human fetal umbilical cord blood (FUCB-MSCs) to culture then label enhanced green fluorescent protein (EGFP) , and to observe skin repair effects of FUCB-MSCs by GFP tracing after exogenous FUCB-MSCs transplantation on to scald wound models of SCID mice.Methods FUCB-MSCs were labeled GFP by transfection with the recombinant retrovirus containing EGFP gen;The established SCID scald mice model were randomLy divided into 3groups, low dose group, high dose group and control group, 6rats each group, 2wounds each mouse, 12wounds in total, then were tail intravenous injected into 0.2mL 1×106, 0.2mL 2×106 GFP-FUCB-MSCs cells, and same volume of medium respectively.On 9days after transplantation, the sections from scald wound area were observed the expression of GFP under the fluorescence microscope and the others were analyzed by the bright-field microscopy after HE staining, and the area of wound surface and the number of wound cells were compared simultaneously.Results After 48h, expression of EGFP in FUCB-MSCs can be seen under the fluorescence microscope, positive rate of GFP was>80％, and after 6weeks GFP expression is still stable, besides, the positive expression of human GFP can be observed after transplantation and there were no fluorescence decay in transplantation after 3weeks.Compared with the control group, there was a significant difference in wound area and wound cell number in the low and high-dose group (P<0.05) .ConclusionGFP can be used as a tracking marker to label FUCB-MSCs during transplantation treatment.It indicates that exogenous FUCB-MSCs can migrate to the scalded wounds via blood circulation system and continuously participate in the repair through SCID mouse.
