本文選題：肉芽組織 + 成纖維細(xì)胞��； 參考：《遵義醫(yī)學(xué)院》2013年碩士論文

【摘要】：目的：觀察體外培養(yǎng)人創(chuàng)面肉芽組織成纖維細(xì)胞的增殖活力、CD分子表型、部分因子及相關(guān)蛋白表達(dá)和體外誘導(dǎo)分化能力等生物學(xué)特性,探討成纖維細(xì)胞在創(chuàng)面愈合早期中的作用,為進(jìn)一步研究提供研究資料。 方法：成人深度燒傷后14天,整塊切取創(chuàng)面肉芽組織,采用機(jī)械法和酶消化法相結(jié)合分離、培養(yǎng)人創(chuàng)面肉芽組織成纖維細(xì)胞,倒置相差顯微鏡下觀察細(xì)胞形態(tài)及增殖情況,計(jì)算原代及第3代細(xì)胞倍增時(shí)間,繪制原代及第3代細(xì)胞生長曲線。應(yīng)用流式細(xì)胞儀檢測原代及第3代細(xì)胞CD105、CD73、CD90及CD44等間充質(zhì)干細(xì)胞表面標(biāo)志,CD34、CD45、CD19、CDllb及HLA-DR等造血干細(xì)胞表面標(biāo)志的表達(dá)情況。取第3代細(xì)胞行免疫細(xì)胞化學(xué)檢測波形蛋白(Vimentin)、角蛋白19(CK19)、CD31和第Ⅷ因子相關(guān)抗原(FⅧ)的表達(dá)情況。并對第3代細(xì)胞進(jìn)行成脂、成骨及成軟骨細(xì)胞多向誘導(dǎo)分化能力的鑒定。 結(jié)果：原代培養(yǎng)的成纖維細(xì)胞呈短梭形、多角形或不規(guī)則形；體外培養(yǎng)連續(xù)多次傳代后細(xì)胞形態(tài)主要以長梭形為主,排列規(guī)則,呈平行狀、放射狀、旋渦狀生長；細(xì)胞增殖良好,有明顯的對數(shù)生長期,體外培養(yǎng)第1-4天,原代及第三代細(xì)胞增殖能力無差異(P0.05),從第5天開始第三代比原代細(xì)胞增殖能力強(qiáng)(P0.05)。人創(chuàng)面肉芽組織成纖維細(xì)胞原代細(xì)胞的倍增時(shí)間為18.7h,第3代細(xì)胞的倍增時(shí)間為15.3h。流式細(xì)胞儀檢測原代及第3代細(xì)胞高表達(dá)CD105、CD73、CD90及CD44等間充質(zhì)干細(xì)胞表面標(biāo)志,不表達(dá)CD34、CD45、CD19、CDllb及HLA-DR等造血干細(xì)胞表面標(biāo)志,且第3代細(xì)胞表達(dá)間充質(zhì)干細(xì)胞表面標(biāo)志比例明顯增高。免疫細(xì)胞化學(xué)檢測波形蛋白、內(nèi)皮細(xì)胞標(biāo)志物CD31陽性表達(dá)；不表達(dá)第Ⅷ因子相關(guān)抗原和角蛋白19。多向誘導(dǎo)分化實(shí)驗(yàn)顯示可向成骨細(xì)胞、軟骨細(xì)胞和脂肪細(xì)胞分化,具有多向分化潛能。 結(jié)論：1、體外培養(yǎng)人創(chuàng)面肉芽組織成纖維細(xì)胞表達(dá)間充質(zhì)干細(xì)胞樣細(xì)胞的CD分子表型,且具有多向分化潛能,該細(xì)胞具有間充質(zhì)干細(xì)胞樣生物學(xué)特性。 2、體外培養(yǎng)人創(chuàng)面肉芽組織成纖維細(xì)胞表達(dá)內(nèi)皮細(xì)胞部分生物學(xué)標(biāo)志物,創(chuàng)面血管內(nèi)皮細(xì)胞通過內(nèi)皮向間質(zhì)轉(zhuǎn)化(EndMT)可能是人創(chuàng)面愈合早期肉芽組織成纖維細(xì)胞的來源之一。
[Abstract]:Objective: to observe the biological characteristics of proliferative activity of fibroblasts from human wound granulation tissue in vitro, such as phenotype of CD, expression of some factors and related proteins, and ability of inducing differentiation in vitro. To investigate the role of fibroblasts in early wound healing, and to provide data for further study. Methods: after 14 days of adult deep burn, the granulation tissue of the wound was cut and separated by mechanical method and enzyme digestion method. The fibroblasts of human wound granulation tissue were cultured. The morphology and proliferation of the granulation tissue were observed under inverted phase contrast microscope. The doubling time of primary and third generation cells was calculated and the growth curves of primary and third generation cells were plotted. Flow cytometry was used to detect the expression of surface markers of primary and third generation cells, such as CD105, CD73, CD90 and CD44, on the surface markers of hematopoietic stem cells, such as CD34, CD45, CD19, CDllb and HLA-DR. The expression of vimentin (Vimentinus), keratin 19 (CK19) CD31 and factor 鈪，

本文編號：1920378