本文選題：人類(lèi)視網(wǎng)膜色素上皮 + 細(xì)胞增殖��； 參考：《福建醫(yī)科大學(xué)》2010年碩士論文

【摘要】： 目的:觀察堿性成纖維細(xì)胞生長(zhǎng)因子(bFGF)、表皮生長(zhǎng)因子(EGF)對(duì)體外人類(lèi)視網(wǎng)膜色素上皮(hRPE)細(xì)胞增殖調(diào)控的劑量-效應(yīng)關(guān)系、時(shí)間-效應(yīng)關(guān)系以及bFGF和EGF的促增殖效應(yīng),探索體外培養(yǎng)hRPE細(xì)胞增殖的最佳培養(yǎng)濃度、作用時(shí)間以及效應(yīng)較強(qiáng)的生長(zhǎng)因子,為hRPE細(xì)胞替代療法治療視網(wǎng)膜變性疾病提供新思路。 方法:用RPE65免疫細(xì)胞化學(xué)染色(SP二步法)鑒定確認(rèn)冷凍、復(fù)蘇的細(xì)胞為hRPE細(xì)胞;用CCK-8法檢測(cè)不同濃度的bFGF及EGF (0,2,4,8,16,32ng/ml)在不同作用時(shí)間(0,24,48,72,96h)對(duì)hRPE增殖的影響。 結(jié)果:在相同培養(yǎng)時(shí)間、不同濃度bFGF和EGF作用下,hRPE細(xì)胞吸光度(OD值)隨bFGF和EGF的濃度增加而增加,且與對(duì)照組比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。在培養(yǎng)相同時(shí)間下,bFGF濃度16ng/ml以及EGF濃度8ng/ml的相鄰濃度實(shí)驗(yàn)組的兩組間比較差異有統(tǒng)計(jì)學(xué)意義(P0.05);bFGF濃度≥16ng/ml以上以及EGF濃度≥8ng/ml的相鄰濃度實(shí)驗(yàn)組兩組間比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。在相同濃度的bFGF或EGF作用下,hRPE細(xì)胞吸光值(OD值)隨時(shí)間增加而增加,且與對(duì)照組比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05);EGF作用時(shí)間在24h以上比相應(yīng)的bFGF實(shí)驗(yàn)組的效應(yīng)強(qiáng),兩組差異有統(tǒng)計(jì)學(xué)意義(P0.05)。在培養(yǎng)相同時(shí)間下,濃度≥4ng/ml的EGF實(shí)驗(yàn)組促增殖效應(yīng)比相應(yīng)的bFGF實(shí)驗(yàn)組強(qiáng),兩組差異有統(tǒng)計(jì)學(xué)意義(P0.05),但濃度為2ng/ml的EGF和bFGF促增殖效應(yīng)的差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論:bFGF和EGF對(duì)體外培養(yǎng)hRPE細(xì)胞增生的調(diào)控都存在一定的劑量-效應(yīng)關(guān)系、時(shí)間-效應(yīng)關(guān)系。EGF對(duì)體外培養(yǎng)hRPE的促增殖效應(yīng)強(qiáng)于bFGF。bFGF的促進(jìn)作用在16ng/ml以上逐漸達(dá)到飽和;EGF的促進(jìn)作用在8ng/ml以上逐漸達(dá)到飽和。研究結(jié)果提示:促體外培養(yǎng)hRPE細(xì)胞增殖的bFGF濃度應(yīng)以16ng/ml、EGF濃度應(yīng)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(hào)：1902701