不同浓度的bFGF和EGF对体外培养的视网膜色素上皮细胞增殖的影响

发布时间：2018-05-17 20:13

  本文选题：人类视网膜色素上皮 + 细胞增殖　； 参考：《福建医科大学》2010年硕士论文

【摘要】： 目的:观察碱性成纤维细胞生长因子(bFGF)、表皮生长因子(EGF)对体外人类视网膜色素上皮(hRPE)细胞增殖调控的剂量-效应关系、时间-效应关系以及bFGF和EGF的促增殖效应,探索体外培养hRPE细胞增殖的最佳培养浓度、作用时间以及效应较强的生长因子,为hRPE细胞替代疗法治疗视网膜变性疾病提供新思路。 方法:用RPE65免疫细胞化学染色(SP二步法)鉴定确认冷冻、复苏的细胞为hRPE细胞;用CCK-8法检测不同浓度的bFGF及EGF (0,2,4,8,16,32ng/ml)在不同作用时间(0,24,48,72,96h)对hRPE增殖的影响。 结果:在相同培养时间、不同浓度bFGF和EGF作用下,hRPE细胞吸光度(OD值)随bFGF和EGF的浓度增加而增加,且与对照组比较差异均有统计学意义(P0.05)。在培养相同时间下,bFGF浓度16ng/ml以及EGF浓度8ng/ml的相邻浓度实验组的两组间比较差异有统计学意义(P0.05);bFGF浓度≥16ng/ml以上以及EGF浓度≥8ng/ml的相邻浓度实验组两组间比较差异无统计学意义(P0.05)。在相同浓度的bFGF或EGF作用下,hRPE细胞吸光值(OD值)随时间增加而增加,且与对照组比较差异均有统计学意义(P0.05);EGF作用时间在24h以上比相应的bFGF实验组的效应强,两组差异有统计学意义(P0.05)。在培养相同时间下,浓度≥4ng/ml的EGF实验组促增殖效应比相应的bFGF实验组强,两组差异有统计学意义(P0.05),但浓度为2ng/ml的EGF和bFGF促增殖效应的差异无统计学意义(P0.05)。 结论:bFGF和EGF对体外培养hRPE细胞增生的调控都存在一定的剂量-效应关系、时间-效应关系。EGF对体外培养hRPE的促增殖效应强于bFGF。bFGF的促进作用在16ng/ml以上逐渐达到饱和;EGF的促进作用在8ng/ml以上逐渐达到饱和。研究结果提示:促体外培养hRPE细胞增殖的bFGF浓度应以16ng/ml、EGF浓度应以8ng/ml为宜。
[Abstract]:Aim: to observe the dose-effect relationship, time-effect relationship and the proliferative effect of bFGF and EGF on the proliferation of human retinal pigment epithelium (RPE) cells induced by basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) in vitro. In order to provide a new idea for the treatment of retinal degeneration by hRPE cell replacement therapy, the best culture concentration, time of action and strong effective growth factor of hRPE cell proliferation in vitro were explored. Methods: the frozen and resuscitated cells were identified as hRPE cells by RPE65 immunocytochemical staining (SP two-step method), and the effects of different concentrations of bFGF and 816ng / ml of EGF on the proliferation of hRPE at different time were detected by CCK-8 assay. Results: at the same culture time, the absorbance and OD value of hRPE cells increased with the increase of bFGF and EGF concentration under different concentrations of bFGF and EGF, and the difference was statistically significant compared with the control group (P 0.05). There were significant differences between the two groups in the concentration of 16ng/ml and 8ng/ml in EGF at the same time. There was no significant difference between the two groups in the concentration of bFGF 鈮，

本文编号：1902701