【摘要】：本研究旨在建立一種簡(jiǎn)單易行、獲取細(xì)胞純度高的絨山羊附睪上皮細(xì)胞體外培養(yǎng)體系。10~12月齡絨山羊附睪頭部分別采用酶消化法和改良組織塊消化法進(jìn)行原代培養(yǎng),利用免疫細(xì)胞化學(xué)染色法和免疫熒光化學(xué)法對(duì)細(xì)胞進(jìn)行鑒定,同時(shí)利用流式細(xì)胞儀和CCK法分別檢測(cè)細(xì)胞純度和增殖情況,在電鏡下觀察細(xì)胞超微結(jié)構(gòu)。結(jié)果表明,兩種培養(yǎng)方法所獲的細(xì)胞均呈島嶼狀克隆生長(zhǎng)和較好的增殖能力,具有活躍的分泌和代謝功能;上皮細(xì)胞特異性的角蛋白18和附睪特異性的谷胱甘肽過氧化物酶5(GPx5)的表達(dá)以及流式細(xì)胞儀結(jié)果顯示,兩種方法所獲細(xì)胞均為純度較高的附睪上皮細(xì)胞,但改良組織塊消化法獲得原代細(xì)胞需要的時(shí)間長(zhǎng),酶消化法獲得原代細(xì)胞時(shí)間短,獲得的細(xì)胞量大。本研究結(jié)果為進(jìn)一步研究絨山羊附睪上皮功能提供了良好的基礎(chǔ)。
[Abstract]:The aim of this study was to establish an in vitro culture system for epididymal epithelial cells of cashmere goats with high cell purity. The epididymal head of 10-12 month-old cashmere goats was primarily cultured by enzyme digestion method and modified tissue block digestion method, respectively. The cell purity and proliferation were detected by flow cytometry and CCK method respectively. The ultrastructure of the cells was observed under electron microscope. The results showed that the cells obtained by the two methods showed island-like clone growth and good proliferation ability, and had active secretory and metabolic functions. The expression of epithelial cell-specific keratin 18 and epididymis-specific glutathione peroxidase 5 (GPx5) and the results of flow cytometry showed that the cells obtained by both methods were high purity epididymal epithelial cells. However, it takes a long time to obtain primary cells by modified tissue block digestion method, and short time to obtain primary cells by enzyme digestion method, and the amount of cells obtained is large. The results of this study provide a good basis for further study of epididymal epithelial function in cashmere goats.
【作者單位】： 內(nèi)蒙古農(nóng)業(yè)大學(xué)動(dòng)物科學(xué)學(xué)院內(nèi)蒙古自治區(qū)動(dòng)物遺傳育種與繁殖重點(diǎn)實(shí)驗(yàn)室;內(nèi)蒙古醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院;
【基金】：國(guó)家自然科學(xué)基金(31460598) 海南省重大科技計(jì)劃(ZDKJ2016017-02)
【分類號(hào)】：S827

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本文編號(hào)：2469339