CM-DiI labeling human bone marrow mesenchymal stem cells and was used in identifying BNDF expressing cell after transplantation

doi:10.3969/j.issn.1006-7795.2017.03.019

Abstract

Abstract: Objective To observe whether the CM-DiI carried by human bone marrow mesenchymal stem cells (BM-MSC) after labeling will contaminate the surrounding cells in vitro and in vivo and be used in identifing brain derived neurotrophic factor (BDNF) production. Methods The human bone marrow derived mesenchymal stem cells were labeled by CM-DiI at different concentrations and the labeling percentages were observed. The CM-DiI labeled human BM-MSCs were co-cultured with green fluorescence protein (GFP) labeled 293T cell for 7 days. Then the contamination of 293T cells by CM-DiI was observed. The cerebral ischemia rat model received CM-DiI labeled human BM-MSCs intravenously transplantation, the overlapping of red fluorescence with host neurons and brain derived neurotrophic factor BDNF in the host brain were observed. Results The human BM-MSCs could be successfully labeled by CM-DiI at 1 000, 200,100, and 20 nmol/L. The concentrations of 1 000 and 200 nmol/L reached more than 98% labeling percentages which were significantly higher than that labeled by 100 and 20 nmol/L(P<0.01). After 7 days co-culture of GFP labeled 293T cells, the CM-DiI labeled human BM-MSCs didn't contaminate the 293T cells. Three days after transplantation, the CM-DiI labeled human BM-MSCs migrated into the infarct area of brain ischemia rats, but didn't bring any red fluorescence into the host cells. CM-DiI labeled human BM-MSCs could be double labeled by BDNF. Conclusion CM-DiI could label human BM-MSCs effectively. The 200 nmol/L CM-DiI labeled human BM-MSCs will not contaminate the surrounding cells both in vitro and in vivo. The CM-DiI (200 nmol/L) labeling can be used in identifying transplanted BM-MSCs which are expressing BDNF.

Key words: mesenchymal stem cells, CM-DiI, transplantation, cerebral infarct, brain derived neurotrophic factor

CLC Number:

Zhao Chunsong, Zou Haiqiang, Li Xiaobo, Yan Xiaoming, Guan Yunqian, Zhang Yu. CM-DiI labeling human bone marrow mesenchymal stem cells and was used in identifying BNDF expressing cell after transplantation[J]. Journal of Capital Medical University, 2017, 38(3): 423-430.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2017.03.019

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2017/V38/I3/423

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