论文部分内容阅读
目的评价骨髓间充质干细胞(MSCs)介导大鼠β-半乳糖苷酶(Lac-Z)基因肺内转移的可行性。方法对原代培养的Lewis大鼠MSCs进行Lac-Z表达载体(pSV-β-Gal)转染和DAPI荧光染色标记,再将MSCs(5×105细胞/每只大鼠)经颈静脉输入同基因背景受体Lewis鼠体内(n=10),48 h 和8周后处死大鼠(每时点5只大鼠),制作肺、脾、肝、肾、骨骼肌标本,荧光显微镜观察荧光标记的移植细胞,将肺组织与X-gal色素原一起孵育检测Lae-Z基因表达产物。结果静脉输入细胞后48 h及 8周,脾、肝、肾组织仅见少量DAPI标记的阳性细胞,骨骼肌组织未见阳性细胞,但肺组织内可见较多 DAPI标记的阳性细胞,主要分布于肺间质和肺泡腔内。经颈静脉输入转染了pSV-β-Gal的MSCs后48 h及8周,肺组织标本与X-gal色素原溶液一起孵育后,显微镜下可见大鼠肺内散布着许多深蓝色标记的MSCs,而脾、肾中则仅见少量深蓝色标记的MSCs,肝、骨骼肌未见深蓝色标记的MSCs。结论颈外静脉注射基因修饰的MSCs具有较好的肺内靶向性,且在受体鼠肺可较长时间表达外源基因。
Objective To evaluate the feasibility of bone marrow mesenchymal stem cells (MSCs) mediated lung metastasis of rat β-galactosidase (Lac-Z) gene. Methods Primary cultured Lewis rat MSCs were transfected with Lac-Z expression vector and DAPI fluorescent staining. MSCs (5 × 10 5 cells / rat) were injected into the same vein through the jugular vein Rats were sacrificed at 48 h and 8 weeks after transplantation (n = 10). Five rats at each time point were used to make lung, spleen, liver, kidney and skeletal muscle samples. Fluorescent microscope Of the transplanted cells, the lung tissue was incubated with X-gal pro-pigment to detect Lae-Z gene expression products. Results At 48 h and 8 weeks after intravenous injection, only a few DAPI positive cells were detected in spleen, liver and kidney, but no positive cells in skeletal muscle. However, more DAPI positive cells were found in lung tissues, mainly in lung Interstitial and alveolar cavity. After 48 h and 8 weeks of transfection of MSCs transfected with pSV-β-Gal through the jugular vein, lung tissue samples were incubated with X-gal proanthocyanidin solution. Microscopically, many dark blue labeled MSCs were found in the lungs of rats , While only a few dark blue labeled MSCs were seen in the spleen and kidney, while no dark blue labeled MSCs were found in liver and skeletal muscle. Conclusion The exogenous jugular vein injection of genetically modified MSCs has good lung targeting and the exogenous gene can be expressed for a long time in the recipient rat lung.