| 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). |
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| 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 |
