云南半细毛羊毛囊干细胞系的建立

发布时间：2018-03-27 15:35

本文选题：云南半细毛羊　切入点：毛囊干细胞　出处：《西北农林科技大学学报(自然科学版)》2017年04期

【摘要】：【目的】研究云南半细毛羊毛囊干细胞(Hair follicle stem cells,HFSCs)的分离培养方法。【方法】用组织块法原代培养云南半细毛羊HFSCs,并进行细胞纯化和传代培养,培养液为无血清培养液DMEM/F12+40ng/mL bFGF+20ng/mL EGF+20μL/mL B27+100IU/mL青霉素+100IU/mL链霉素。对建立的细胞系进行免疫组化、RT-PCR和流式细胞术鉴定。【结果】HFSCs体积小,为贴壁生长细胞,呈典型的铺路石状,核质比高,在倒置显微镜下胞体透亮、折光性强。RT-PCR分析表明,HFSCs表达K14、K19和β1-integrin,但是不表达CD271和nestin。免疫组化分析结果表明,HFSCs表达K14,不表达CD271。流式细胞术鉴定表明,HFSCs表达K14,阳性率为93.5%;不表达CD271。【结论】建立了云南半细毛羊HSFCs无血清体外培养技术体系。
[Abstract]:[objective] to study the method of isolation and culture of hair follicle stem cells (Hair follicle stem cells) of Yunnan semi-fine wool sheep. [methods] the primary culture of Yunnan semi-fine wool sheep was carried out by tissue block method, and the cells were purified and subcultured. The culture medium was DMEM/F12 40ng/mL bFGF 20ng/mL EGF 20 渭 L/mL B27 100IU/mL penicillin 100IU/mL streptomycin. The established cell lines were identified by immunohistochemistry RT-PCR and flow cytometry. [results] HFSCs was a small, adherent growth cell, and had typical paving stone shape. The ratio of nucleus to cytoplasm is high, and the cell body is bright under inverted microscope. The results of immunocytochemical analysis showed that HSCs expressed K14 K14 and 尾 1-integrin, but not CD271 and Neisin. The results of immunohistochemical analysis showed that HSCs expressed K14 and CD271. Flow cytometry showed that HSCs expressed K14 with a positive rate of 93.50.Conclusion the expression of CD271was established. HSFCs serum-free culture technique system of South semi-fine wool sheep in vitro.
【作者单位】：云南省畜牧兽医科学院;
【基金】：国家现代农业产业技术体系建设项目(CARS-40) 云南省应用基础研究青年项目(2012FD083)
【分类号】：S826

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