本文選題：雞 + 肌腱干細胞�。� 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胞的分離培養(yǎng)體系和鑒定方法,試驗使用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胞,用結(jié)晶紫染色細胞克隆團塊,選擇50,500,5 000個/cm~2密度接種7~10 d,鏡下觀察細胞形態(tài),用細胞計數(shù)儀測定細胞增殖能力,用分化誘導試劑對細胞進行成脂與成骨分化誘導,采用油紅O和堿性磷酸酶進行染色鑒定。結(jié)果表明:原代細胞及傳代細胞為長梭形,接種7~10 d后開始形成克隆團塊,細胞生長曲線呈S型。500個/cm~2的細胞密度接種細胞克隆能力與增殖能力最強;將該密度接種細胞消化后,按1×10~4個/cm~2接種,分別對其進行成脂與成骨誘導,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胞,克隆增殖能力強,且具有分化多種細胞的潛能。
[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】： 南通大學比較醫(yī)學研究所;南通大學神經(jīng)再生重點實驗室;南通大學附屬醫(yī)院;
【基金】：國家自然科學基金項目(81401797) 南通大學研究生科技創(chuàng)新計劃項目(YKC16042;YKC16061)
【分類號】：Q813.1;R687.2

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