本文選題：雞 + 肌腱干細(xì)胞　； 參考：《黑龍江畜牧獸醫(yī)》2017年19期

【摘要】：為了建立雞肌腱干細(xì)胞的分離培養(yǎng)體系和鑒定方法,試驗(yàn)使用Ⅰ型膠原酶消化20日齡雞胚肌腱組織,低密度接種得到雞肌腱干細(xì)胞,用結(jié)晶紫染色細(xì)胞克隆團(tuán)塊,選擇50,500,5 000個(gè)/cm~2密度接種7~10 d,鏡下觀察細(xì)胞形態(tài),用細(xì)胞計(jì)數(shù)儀測定細(xì)胞增殖能力,用分化誘導(dǎo)試劑對細(xì)胞進(jìn)行成脂與成骨分化誘導(dǎo),采用油紅O和堿性磷酸酶進(jìn)行染色鑒定。結(jié)果表明:原代細(xì)胞及傳代細(xì)胞為長梭形,接種7~10 d后開始形成克隆團(tuán)塊,細(xì)胞生長曲線呈S型。500個(gè)/cm~2的細(xì)胞密度接種細(xì)胞克隆能力與增殖能力最強(qiáng);將該密度接種細(xì)胞消化后,按1×10~4個(gè)/cm~2接種,分別對其進(jìn)行成脂與成骨誘導(dǎo),可發(fā)現(xiàn)成脂分化過程中有脂滴形成,油紅O染色呈陽性;成骨分化過程中有鈣結(jié)節(jié)形成,堿性磷酸酯酶染色呈陽性。說明消化20日齡雞胚肌腱組織可成功分離出雞肌腱干細(xì)胞,克隆增殖能力強(qiáng),且具有分化多種細(xì)胞的潛能。
[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.
【作者單位】： 南通大學(xué)比較醫(yī)學(xué)研究所;南通大學(xué)神經(jīng)再生重點(diǎn)實(shí)驗(yàn)室;南通大學(xué)附屬醫(yī)院;
【基金】：國家自然科學(xué)基金項(xiàng)目(81401797) 南通大學(xué)研究生科技創(chuàng)新計(jì)劃項(xiàng)目(YKC16042;YKC16061)
【分類號】：Q813.1;R687.2

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