大鼠骨髓内皮祖细胞移植到裸鼠体内的分化

    Differentiation of rat marrow-derived endothelial progenitor cells transplanted into nude mice

    • 摘要: 目的:研究骨髓内皮祖细胞(EPCs)在裸鼠体内分化情况,进一步揭示其生物学特性。方法:密度梯度离心法分离大鼠骨髓EPCs;Hoechst33342标记原代(第10天)的EPCs注射到裸鼠背部两侧皮下,观测注射部位的变化及生成物的大小;移植第50天,剥离EPCs形成的皮下肿物,HE及免疫荧光染色,检测细胞CD31、Flk1、CD34的表达情况。结果:裸鼠皮下接种EPCs第20天,形成(0.13±0.018)cm2球形皮下肿物,逐渐增大,第40天开始处于生长平台期;HE染色组织内有管腔样结构、肌肉样组织及脂肪样细胞,同视野荧光显微镜下可见Hoechst标记的细胞;免疫荧光检测显示,Hoechst标记细胞抗原CD31、Flk1、CD34表达阳性。结论:大鼠骨髓EPCs移植到裸鼠体内一部分分化为内皮样细胞,并形成管腔样结构,另一部分分化为非内皮系细胞。

       

      Abstract: Objective: To study on differentiation of marrow-derived endothelial progenitor cells(EPCs) in vivo,and to further reveal EPCs' biological characteristics.Methods: Rat bone marrow-derived EPCs were separated by density gradient centrifugation,and cultrued in vitro;and then labelled with Hoechst 33342 and collected.The labelled EPCs were injected subcutaneously into nude mice's back,on the both sides of the spinal.Animals were observed daily and measured the size of the subcutaneous masses.At the 50th day after transplantation,the subcutaneous masses were removed and then performed HE and immunofluorescent staining to detect the expression of antigens CD31,Flk1,CD34.Results: At the 20th day after transplantation,we observed the subcutaneous masses with size of(0.13±0.018) cm2 on the nude mice's back,and then the subcutaneous masses were gradually increasing.At the 40th day,EPCs began to get into period of growth stagnation.Tube-like structure,muscle tissue and fatty-like cells were observed through HE staining,at the same vision,blue fluorescent-marked cells were observed through fluorescence microscopy.The result of Immunofluorescence staining showed that some of the blue fluorescent cells expressed antigens CD31,Flk1,CD34.Conclusions: The part of the EPCs transplanted to nude mice differentiated into endothelial-like cells and formed tube-like structure,but the other part differentiated into non-endothelial cells.

       

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