本文選題：人類視網膜色素上皮 + 細胞增殖��； 參考：《福建醫(yī)科大學》2010年碩士論文

【摘要】： 目的:觀察堿性成纖維細胞生長因子(bFGF)、表皮生長因子(EGF)對體外人類視網膜色素上皮(hRPE)細胞增殖調控的劑量-效應關系、時間-效應關系以及bFGF和EGF的促增殖效應,探索體外培養(yǎng)hRPE細胞增殖的最佳培養(yǎng)濃度、作用時間以及效應較強的生長因子,為hRPE細胞替代療法治療視網膜變性疾病提供新思路。 方法:用RPE65免疫細胞化學染色(SP二步法)鑒定確認冷凍、復蘇的細胞為hRPE細胞;用CCK-8法檢測不同濃度的bFGF及EGF (0,2,4,8,16,32ng/ml)在不同作用時間(0,24,48,72,96h)對hRPE增殖的影響。 結果:在相同培養(yǎng)時間、不同濃度bFGF和EGF作用下,hRPE細胞吸光度(OD值)隨bFGF和EGF的濃度增加而增加,且與對照組比較差異均有統(tǒng)計學意義(P0.05)。在培養(yǎng)相同時間下,bFGF濃度16ng/ml以及EGF濃度8ng/ml的相鄰濃度實驗組的兩組間比較差異有統(tǒng)計學意義(P0.05);bFGF濃度≥16ng/ml以上以及EGF濃度≥8ng/ml的相鄰濃度實驗組兩組間比較差異無統(tǒng)計學意義(P0.05)。在相同濃度的bFGF或EGF作用下,hRPE細胞吸光值(OD值)隨時間增加而增加,且與對照組比較差異均有統(tǒng)計學意義(P0.05);EGF作用時間在24h以上比相應的bFGF實驗組的效應強,兩組差異有統(tǒng)計學意義(P0.05)。在培養(yǎng)相同時間下,濃度≥4ng/ml的EGF實驗組促增殖效應比相應的bFGF實驗組強,兩組差異有統(tǒng)計學意義(P0.05),但濃度為2ng/ml的EGF和bFGF促增殖效應的差異無統(tǒng)計學意義(P0.05)。 結論:bFGF和EGF對體外培養(yǎng)hRPE細胞增生的調控都存在一定的劑量-效應關系、時間-效應關系。EGF對體外培養(yǎng)hRPE的促增殖效應強于bFGF。bFGF的促進作用在16ng/ml以上逐漸達到飽和;EGF的促進作用在8ng/ml以上逐漸達到飽和。研究結果提示:促體外培養(yǎng)hRPE細胞增殖的bFGF濃度應以16ng/ml、EGF濃度應以8ng/ml為宜。
[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，

本文編號：1902701