發(fā)布時(shí)間：2018-07-06 10:37

  本文選題：細(xì)胞培養(yǎng) + ARPE-19細(xì)胞��； 參考：《重慶醫(yī)科大學(xué)》2011年碩士論文

【摘要】：目的1掌握人視網(wǎng)膜色素上皮細(xì)胞系A(chǔ)RPE-19細(xì)胞培養(yǎng)、傳代和凍存技術(shù)。2利用免疫熒光技術(shù)探討S100A7蛋白在體外培養(yǎng)人視網(wǎng)膜色素上皮細(xì)胞(retinal pigment epithelial cell, RPE)中的表達(dá)及其分布狀況,并聯(lián)合應(yīng)用Western blot技術(shù)檢測(cè)S100A7蛋白在該細(xì)胞的表達(dá)。3初步探討S100A7在ARPE-19細(xì)胞增殖中的作用,為相關(guān)疾病的診斷治療提供新的思路。 方法1利用細(xì)胞培養(yǎng)方法傳代培養(yǎng)ARPE-19細(xì)胞。2取第3～6代ARPE-19細(xì)胞,運(yùn)用免疫熒光技術(shù)檢測(cè)S100A7蛋白在ARPE-19細(xì)胞中的表達(dá)及其分布情況。3提取ARPE-19總蛋白,運(yùn)用Western blot技術(shù)檢測(cè)S100A7蛋白在ARPE-19細(xì)胞的表達(dá)。4將ARPE-19細(xì)胞接種在96孔細(xì)胞培養(yǎng)板中,用不同濃度S100A7蛋白抗體處理細(xì)胞,然后運(yùn)用MTT方法檢測(cè)ARPE-19細(xì)胞增殖情況,進(jìn)而初步確定S100A7在ARPE-19細(xì)胞增殖中的作用。 結(jié)果1成功培養(yǎng)ARPE-19細(xì)胞,并且細(xì)胞能夠完全滿足該課題中所有實(shí)驗(yàn)的需要。2免疫熒光方法顯示實(shí)驗(yàn)組中ARPE-19細(xì)胞存在S100A7蛋白的表達(dá),表達(dá)部位為該細(xì)胞的細(xì)胞膜,而在對(duì)照組中沒有該蛋白表達(dá)。3 Western blot技術(shù)進(jìn)一步證實(shí)S100A7蛋白在ARPE-19細(xì)胞中的表達(dá)。4.MTT實(shí)驗(yàn)初步確定S100A7蛋白能夠促進(jìn)ARPE-19細(xì)胞的增殖過程。 結(jié)論本實(shí)驗(yàn)首次發(fā)現(xiàn)并證實(shí)S100A7蛋白在人RPE細(xì)胞中的表達(dá),并且初步確定其在RPE細(xì)胞增殖過程中起促進(jìn)作用,從而為研究人視網(wǎng)膜疾病特別是增殖性玻璃體視網(wǎng)膜疾病、糖尿病視網(wǎng)膜疾病提供了新的方向,為該類疾病的治療提供了新的靶點(diǎn)。
[Abstract]:Objective 1 to investigate the expression and distribution of S100A7 protein in human retinal pigment epithelial cell line ARPE-19 by immunofluorescence technique. The expression of S100A7 protein in ARPE-19 cells was detected by Western blot, and the role of S100A7 protein in the proliferation of ARPE-19 cells was preliminarily studied, which provided a new idea for the diagnosis and treatment of related diseases. Methods 1 ARPE-19 cells were subcultured by cell culture method. ARPE-19 cells were collected from ARPE-19 cells. The expression and distribution of S100A7 protein in ARPE-19 cells were detected by immunofluorescence technique, and the total ARPE-19 protein was extracted from ARPE-19 cells. Western blot was used to detect the expression of S100A7 protein in ARPE-19 cells. ARPE-19 cells were inoculated into 96-well cell culture plates. The cells were treated with different concentrations of S100A7 protein antibody, and then the proliferation of ARPE-19 cells was detected by MTT assay. Furthermore, the role of S100A7 in the proliferation of ARPE-19 cells was preliminarily determined. Results (1) ARPE-19 cells were successfully cultured, and the cells could fully meet the needs of all the experiments in this study. 2 immunofluorescence method showed that there was S100A7 protein expression in ARPE-19 cells in the experimental group, and the expression location was the cell membrane of ARPE-19 cells. However, the expression of S100A7 protein in ARPE-19 cells was not further confirmed by Western blot. 4. The results showed that S100A7 protein could promote the proliferation of ARPE-19 cells. Conclusion the expression of S100A7 protein in human RPE cells was found and confirmed for the first time, and it was preliminarily confirmed that S100A7 protein plays a role in promoting the proliferation of RPE cells, which may be useful for the study of human retinal diseases, especially proliferative vitreoretinopathy. Diabetic retinopathy provides a new direction and a new target for the treatment of diabetic retinopathy.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R774.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Application of Two-Dimensional Electrophoresis in the Research of Retinal Proteins of Diabetic Rat[J];Cellular & Molecular Immunology;2007年01期

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本文編號(hào)：2102505