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微囊化血管内皮细胞生长因子基因修饰NIH3T3细胞的制备及体外培养
韩焱福1,宋建星1*,刘军1*,尹东锋2,陈玉林3,吕川1,杨硕成1
0
(1.第二军医大学长海医院整形外科,上海 200433;2.第二军医大学药学院药剂学教研室,上海 200433;3.第二军医大学长海医院烧伤科,上海 200433)
摘要:
目的:制备微囊化血管内皮细胞生长因子(VEGF)基因修饰NIH3T3细胞,并研究微囊化技术对VEGF基因修饰NIH3T3细胞增殖及其分泌VEGF功能的影响。方法:应用海藻酸钠-氯化钡技术制备微囊化VEGF基因修饰NIH3T3细胞,并与未微囊化VEGF基因修饰NIH3T3细胞对照培养,在倒置相差显微镜下观察微囊及细胞形态,用MTT法和PI染色流式细胞术检测细胞的增殖及活性情况。每48 h收集微囊化及未微囊化基因修饰NIH3T3细胞培养上清,-20℃保存,ELISA法检测培养上清VEGF的含量。结果:微囊形态较圆整,其内细胞生长良好;两组细胞增殖与活性及培养上清中VEGF含量无统计学差异。结论:微囊化并不影响基因修饰细胞的生长代谢功能,与对照组比较,在体外培养时生物学特性无明显差异,为研究微囊化基因修饰细胞体内移植奠定了基础。
关键词:  血管内皮细胞生长因子  NIH3T3细胞  微囊  细胞培养
DOI:10.3724/SP.J.1008.2008.00491
投稿时间:2007-12-21修订日期:2008-02-27
基金项目:上海市卫生局青年科研基金(2006Y41).
Preparation and culture of NIH3T3 cells harboring microencapsulated VEGF gene
HAN Yan-fu1,SONG Jian-xing1*,LIU Jun1*,YIN Dong-feng2,CHEN Yu-lin3,Lv Chuan1,YANG Shuo-cheng1
(1.Department of Plastic Surgery,Changhai Hospital,Second Military Medical University,Shanghai 200433,China;2.Department of Pharmaceutics,School of Pharmacy,Second Military Medical University,Shanghai 200433;3.Department of Burns,Changhai Hospital,Second Military Medical University,Shanghai 200433)
Abstract:
Objective:To prepare NIH3T3 cells harboring microencapsulated VEGF gene and investigate the proliferation,activity and metabolic function of the modified cells.Methods: Microencapsulated VEGF modified NIH3T3 cells were prepared through an alginate-BaCl2 process.Morphological appearance of the microencapsulation and the cell morphology were observed under inverted phase microscope; untreated NIH3T3 cells served as control.The concentrations of VEGF in the culture supernatant (collected every 48 hours) were measured by ELISA; the proliferation and vitality of the cells were examined by MTT assay and flow cytometry with PI staining.Results: The microcapsules were round in shape and the cells grew well.There was no significant difference in the concentrations of VEGF,MTT values and vitalities of cells between the 2 groups.Conclusion: The growth and metabolic functions of NIH3T3 cells are not influenced by microencapsulated NIH3T3 cells harboring VEGF gene.The bio-properties of modified cells are similar to those of the control cells,which lays a foundation for transplantation of microencapsulated VEGF modified NIH3T3 cells in vivo.
Key words:  vascular endothelial growth factor  NIH3T3 cells  microencapsulation  cell culture