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目的:探讨体外重组基底膜侵袭实验中最佳趋化条件及细胞计数方法.方法:采用体外重组基底膜侵袭实验方法,加入不同趋化 剂:成纤维细胞条件培养液、纤维黏连蛋白(FN)、条件培养液与FN联合;HE染色后,采用40×光镜随机视野计数法及利用显微 照相系统全滤膜计数法计数.结果:趋化剂组穿膜细胞数显著高于对照组;条件培养液和FN联合穿膜细胞数显著高于单独条件 培养液组和FN组;条件培养液组穿膜细胞数高于FN组;随机视野计数法与全滤膜计数法相比所得结果误差大.结论:趋化剂能 显著增加细胞的定向穿膜侵袭能力,不同趋化剂的趋化能力各有不同;全滤膜计数法是一种客观准确的细胞计数法.
OBJECTIVE: To investigate the optimal chemotactic conditions and cell counting methods in in vitro invasion assay of recombinant basement membrane.Methods: In vitro recombinant basement membrane invasion assay was used to add different chemotactic agents: fibroblast conditioned medium, fibronectin (FN ), Conditioned medium and FN combined; HE staining, the use of 40 × light microscope random field counting method and the use of microphotography system of total membrane counting count results: chemotaxis group transmembrane cell number was significantly higher than the control group ; The number of conditioned medium and FN combined with transmembrane cell was significantly higher than that of conditioned medium and FN group; the number of transmembrane cells in conditioned medium group was higher than that of FN group; the difference between random field counting method and total membrane counting method Large.Conclusion: Chemotactic agents can significantly increase the cell penetrating ability, chemotactic ability of different chemotactic agents are different; total membrane counting method is an objective and accurate cell counting method.