PDGF诱导大鼠主动脉平滑肌细胞迁移模型的建立  

Establishment of assay for smooth muscle cell migration induced by PDGF in rat aorta

作　　者：崔颖[1] 白柳[2] 张金超[2] 杨福春[2] 李胜[3] 赵莹[1] 高颖[1,2] 

机构地区：[1]辽宁省医学细胞分子生物学重点实验室,辽宁大连116044 [2]大连医科大学生物化学教研室,辽宁大连116044 [3]大连医科大学附属第一医院神经内科,辽宁大连116011

出　　处：《大连医科大学学报》2009年第3期242-244,252,共4页Journal of Dalian Medical University

基　　金：辽宁省教育厅科技项目(20060196);辽宁省科技厅自然基金项目(20072166)

摘　　要：[目的]建立PDGF诱导的大鼠主动脉平滑肌细胞(A7r5)迁移的模型。[方法]采用改良的Boyden小室法,以不同浓度(1、5、10、20、50和100 ng/mL)PDGF-BB为诱导剂,将不同浓度(1×105/mL、2×105/mL、4×105/mL)的A7r5细胞置入Boyden小室,分别培养5、7、9 h。迁移膜经Giemsa染色后,显微镜下观察迁移到下室的细胞,进行细胞计数。[结果]在无PDGF诱导时,迁移细胞数为每视野10个左右;而以10 ng/mL PDGF-BB诱导时,迁移细胞数每视野可达70个左右;当PDGF浓度达到100 ng/mL时,细胞迁移数降至每视野10个左右。随细胞浓度的增高,迁移细胞数目增加,细胞浓度为1×105/mL时,7 h迁移细胞数目为(48±8.944)个;2×105/mL细胞在5 h左右发生迁移的数目为(55±12.864)个,随时间延长而增多,在7 h前后增幅最明显,达(78±15.828)个(P<0.05);但细胞浓度为4×105/mL时,7 h迁移细胞数为(84±12.213)个(P>0.05)。[结论]以10 ng/mL PDGF-BB为诱导剂,以2×105/mL浓度接种,培养7 h建立的迁移模型为最佳。[ Objective] To establish the model of rat aorta smooth muscle cell（ A7r5 ） migration induced by PDGF. [ Methods]VSMC migration in response to different concentration of PDGF -BB （1,5,10,20,50 and 100 ng/mL） was determined by a modified Boyden chamber method. The various concentration of cell suspension（ 1 ×10^5/mL, 2 ×10^5/mL and 4 ×10^5/mL） was added into the upper chamber, ceils were incubated for 5, 7 and 9 hr respectively. The cells migrated to the lower surface of the filter were stained with giemsa solution, and observed by a light microscope（400）. [ Results] The number of migrated cells （NMC） was about 10/visual field without PDGF. The NMC reached 70/visual field when the cells induced by 10 ng/mL PDGF. The NMC dropped to 10/visual field when the ceils induced by 100 ng/mL PDGF. The NMC with 1 ×10^5/mL concerntration was 48 ±8. 944 at 7 hr. The NMC with 2 ×10^5mL concerntration was 55± 12. 864 at 5 hr, and increased dramatically at 7 hr, the NMC reached 78± 15. 828 （P 〈 0.05）, but only was 84±12. 213 （ P 〉 0. 05 ） when the cell concentration reached 4 ×10^5 /mL. [ Conclusion] The optimal condition is that 2 ×10^5/mL cells are added in the upper chamber and the migrated cells are induced for 7 hr. by using 10 ng/mL PDGF - BB.

关 键 词：平滑肌细胞 BOYDEN小室 细胞迁移 血小板源性生长因子 

分 类 号：Q25[生物学—细胞生物学]