血清含量对HaCaT细胞生长特性及迁移能力的影响

Effect of serum content on the growth characteristic in the HaCaT cells

关键词:  HaCaT细胞  划痕实验  创伤修复  方案优化

Key words:  HaCaT cell  wound scratch assay  wound healing  optimization experiment

摘要:

[目的] 考察不同血清含量以及培养不同时间对人永生化角质形成细胞(HaCaT)细胞的生长及迁移特性的影响,为HaCaT细胞划痕实验条件优化提供实验数据支撑。[方法] 培养HaCaT细胞,按10%血清浓度全培基(全培基阳性对照组)、2%低浓度血清培基(低血清培基组)及无血清培基(对照组)分组,通过进行细胞划痕实验,CCK-8细胞活力检测、Brdu细胞增殖实验,分析HaCaT生长特性。[结果] 在24 h内,低血清培基组HaCaT细胞与全培基组相比,细胞增殖能力无明显差异,细胞活力有增高的趋势。细胞孵育48 h后,全培基组细胞增殖能力及活力明显升高。24~48 h,全培基组细胞增殖能力及细胞活力的提升速度明显高于低血清培基组。[结论] 培养基中血清含量对HaCaT细胞生长特性及迁移能力具有重要影响,培养24 h内低血清培养条件有利于划痕实验的观察与测定。

Abstract:

[Objective] By studying the growth characteristic of the HaCaT cells in conditions of the serum free medium, low serum medium and the serum medium. Put forward the optimization scheme of the HaCaT Wound scratch assay. [Methods] Culture HaCaT cell, grouped into 10% serum medium (positive control group),2% low serum prudential (low serum prudential group) and serum free prudential (control group), through the HaCaT wound scratch assay,CCK-8 assay, Brdu assay, analysis the HaCaT growth characteristics. [Results] With in 24 h, low serum prudential group HaCaT cells compared with the positive control group,no difference between the cell proliferation, cell vitality had increase tendency, incubation cells after 48 h, positive control group cell proliferation ability and cell viability higher than low serum prudential group. Between 24 h and 48 h, 10% serum medium positive control group cell proliferation ability and speed of cell viability was obviously higher than that of low serum prudential group. [Conclusion] If it is only in the actual experiments, study drug in a short period of time (within 24 h) on HaCaT cell migration ability, the influence of culture in serum-free conditions may be more appropriate.