鸡肌腱干细胞的分离培养与初步鉴定

发布时间：2018-04-30 13:51

本文选题：鸡 + 肌腱干细胞　；参考：《黑龙江畜牧兽医》2017年19期

【摘要】：为了建立鸡肌腱干细胞的分离培养体系和鉴定方法,试验使用Ⅰ型胶原酶消化20日龄鸡胚肌腱组织,低密度接种得到鸡肌腱干细胞,用结晶紫染色细胞克隆团块,选择50,500,5 000个/cm~2密度接种7~10 d,镜下观察细胞形态,用细胞计数仪测定细胞增殖能力,用分化诱导试剂对细胞进行成脂与成骨分化诱导,采用油红O和碱性磷酸酶进行染色鉴定。结果表明:原代细胞及传代细胞为长梭形,接种7~10 d后开始形成克隆团块,细胞生长曲线呈S型。500个/cm~2的细胞密度接种细胞克隆能力与增殖能力最强;将该密度接种细胞消化后,按1×10~4个/cm~2接种,分别对其进行成脂与成骨诱导,可发现成脂分化过程中有脂滴形成,油红O染色呈阳性;成骨分化过程中有钙结节形成,碱性磷酸酯酶染色呈阳性。说明消化20日龄鸡胚肌腱组织可成功分离出鸡肌腱干细胞,克隆增殖能力强,且具有分化多种细胞的潜能。
[Abstract]:In order to establish the isolation culture system and identification method of chicken tendon stem cells, the 20 day old chicken embryo tendon tissue was digested with type I collagenase. The chicken tendon stem cells were inoculated with low density, and the colony blocks were stained with crystal violet. Cell morphology was observed under microscope, cell proliferation ability was measured by cell counter, adipogenic and osteogenic differentiation was induced by differentiation induction reagent, oil red O and alkaline phosphatase staining were used to identify the cells. The results showed that the primary cells and the passage cells were long fusiform, and the clones began to form after 7 days of inoculation. The cell growth curve showed S type. 500 / cm ~ (-2) cell density showed the strongest clone ability and proliferation ability of inoculated cells. After the density was digested, the cells were inoculated with 1 脳 10 ~ 4 / cm ~ (-2) to induce adipogenesis and osteogenesis. It was found that lipid droplets were formed in the process of adipogenic differentiation, oil red O staining was positive, and calcium nodules were formed during osteogenic differentiation. Alkaline phosphatase staining was positive. The results showed that chicken tendon stem cells could be isolated successfully by digesting 20 days old chicken embryo tendon tissue, and had the ability to clone and proliferate, and had the potential to differentiate many kinds of cells.
【作者单位】：南通大学比较医学研究所;南通大学神经再生重点实验室;南通大学附属医院;
【基金】：国家自然科学基金项目(81401797) 南通大学研究生科技创新计划项目(YKC16042;YKC16061)
【分类号】：Q813.1;R687.2

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