本文選題：角質(zhì)形成細(xì)胞　切入點(diǎn)：細(xì)胞培養(yǎng)　出處：《華中科技大學(xué)》2011年碩士論文

【摘要】：第一部分人角質(zhì)形成細(xì)胞的原代培養(yǎng) 目的 建立人表皮角質(zhì)形成細(xì)胞的分離和培養(yǎng)方法,為后續(xù)實(shí)驗(yàn)提供基礎(chǔ)。 方法 取青少年包皮環(huán)切術(shù)后的包皮組織,經(jīng)無菌處理后,用常用的胰酶消化法分離表皮和真皮,分離出的表皮細(xì)胞用無血清角質(zhì)形成細(xì)胞培養(yǎng)基培養(yǎng)并用免疫細(xì)胞化學(xué)方法進(jìn)行鑒定。 結(jié)果 用無血清培養(yǎng)基培養(yǎng)的角質(zhì)形成細(xì)胞,成纖維細(xì)胞污染少,貼壁率高,熒光顯微鏡下細(xì)胞胞漿呈角蛋白陽性染色。6代以內(nèi)的細(xì)胞活性均較好。 結(jié)論 用常規(guī)的酶消化法分離表皮,以無血清角質(zhì)形成細(xì)胞培養(yǎng)基培養(yǎng)能成功培養(yǎng)出角質(zhì)形成細(xì)胞。 第二部分TLR2在抗紅色毛癬菌感染中對(duì)角質(zhì)形成細(xì)胞分泌γ-IFN和IL-8的影響 目的 觀察紅色毛癬菌刺激人角質(zhì)形成細(xì)胞后γ-IFN及IL-8的變化,以及TLR-2對(duì)γ-IFN和IL-8的分泌的影響;探討TLR-2在抗皮膚癬菌感染中的作用。 方法 用紅色毛癬菌懸液分別刺激TLR2抗體處理前后的角質(zhì)形成細(xì)胞,采用ELISA方法檢測(cè)不同時(shí)間點(diǎn)細(xì)胞上清液中γ-IFN及IL-8的濃度,并設(shè)置陰性對(duì)照;比較TLR2抗體處理前后γ-IFN及IL-8濃度的變化。 結(jié)果 紅色毛癬菌刺激角質(zhì)形成細(xì)胞后,γ-IFN及IL-8濃度明顯升高(P㩳0.05),γ-IFN在4h即達(dá)到(85.36±4.54)pg/ml,16h后達(dá)到(445.58±13.99)pg/ml ;IL-8在4h即達(dá)到(545.426±39.784)pg/ml,16h后達(dá)到(977.182±55.288)pg/ml;用TLR2抗體中和TLR2后,清液中IL-8的濃度在2h、4h、8h、16h各時(shí)間點(diǎn)較中和前低,差異有統(tǒng)計(jì)學(xué)意義(P㩳0.05);γ-IFN的濃度2h、4h、8h時(shí)間點(diǎn)較中和前低,差異有統(tǒng)計(jì)學(xué)意義(P㩳0.05),而在16h時(shí)間點(diǎn),上清液中γ-IFN的濃度與中和前比較略低,但差異沒有統(tǒng)計(jì)學(xué)意義(P㧐0.05)。 結(jié)論 紅色毛癬菌刺激角質(zhì)形成細(xì)胞后,可促進(jìn)角質(zhì)形成細(xì)胞分泌γ-IFN和IL-8;TLR2在角質(zhì)形成細(xì)胞分泌γ-IFN和IL-8的過程中發(fā)揮重要的調(diào)節(jié)作用。
[Abstract]:Part one primary culture of human keratinocytesPurposeThe method of isolation and culture of human epidermal keratinocytes was established to provide the basis for further experiments.MethodThe tissue of prepuce after circumcision of juvenile prepuce was removed. After sterile treatment, the epidermis and dermis were separated by common trypsin digestion method.The isolated epidermal cells were cultured on serum-free keratinocytes medium and identified by immunocytochemistry.ResultThe keratinocytes cultured in serum-free medium had less contamination and high adhesion rate. The cytoplasm of keratin positive staining could be found in the cytoplasm of the keratin positive cells under fluorescence microscope.ConclusionKeratinocytes were isolated by enzyme digestion and cultured on serum-free keratinocytes medium.The second part: the effect of TLR2 on the secretion of 緯 -IFN and IL-8 by keratinocytes against Trichophyton rubrum infectionPurposeTo observe the changes of 緯 -IFN and IL-8 in human keratinocytes stimulated by Trichophyton rubrum and the effect of TLR-2 on the secretion of 緯 -IFN and IL-8, and to explore the role of TLR-2 in anti-tinea dermatophytes infection.MethodKeratinocytes were stimulated by Trichophyton rubrum suspension before and after TLR2 antibody treatment. The concentrations of 緯 -IFN and IL-8 in supernatant of cells at different time points were detected by ELISA method, and negative control was set.To compare the changes of 緯 -IFN and IL-8 before and after TLR2 antibody treatment.Result綰㈣壊姣涚櫍鑿屽埡嬋，

本文編號(hào)：1705884