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人類軟骨糖蛋白39誘導(dǎo)關(guān)節(jié)軟骨祖母細胞的成軟骨分化

發(fā)布時間:2018-03-05 18:38

  本文選題:軟骨細胞 切入點:糖蛋白類 出處:《中國組織工程研究》2015年42期  論文類型:期刊論文


【摘要】:背景:有研究表明人類軟骨糖蛋白39與骨關(guān)節(jié)軟骨的退變與修復(fù)具有一定的關(guān)系,但其具體的作用機制并不十分明確。目的:觀察人類軟骨糖蛋白39對成人膝關(guān)節(jié)軟骨祖母細胞成軟骨誘導(dǎo)分化的影響。方法:取成人關(guān)節(jié)軟骨,消化分離培養(yǎng)關(guān)節(jié)軟骨祖母細胞;流式細胞儀檢測傳代細胞中能夠表達CD105、CD166的細胞量并進行分離提純。將分離的軟骨祖母細胞采用單層培養(yǎng)法培養(yǎng),傳代培養(yǎng)至第2代后向分離培養(yǎng)所得的軟骨祖母細胞中,分別經(jīng)過含人類軟骨糖蛋白39成軟骨培養(yǎng)基及普通成軟骨誘導(dǎo)培養(yǎng)基的誘導(dǎo)培養(yǎng)14 d后,通過免疫組織化學(xué)染色觀察經(jīng)誘導(dǎo)后細胞中Ⅱ型膠原的表達及通過大體組織學(xué)觀察評估軟骨的形成。結(jié)果與結(jié)論:關(guān)節(jié)軟骨組織中可以分離出能夠表達CD105、CD166的關(guān)節(jié)軟骨祖母細胞,軟骨祖母細胞經(jīng)過誘導(dǎo)分化后逐漸聚集并形成結(jié)節(jié),經(jīng)誘導(dǎo)后Ⅱ型膠原免疫細胞化學(xué)著色陽性,且經(jīng)人類軟骨糖蛋白39誘導(dǎo)細胞形成的結(jié)節(jié)更大,Ⅱ型膠原表達更多。結(jié)果表明,成人關(guān)節(jié)軟骨中能夠分離培養(yǎng)出具有成軟骨分化能力的干細胞系細胞即軟骨祖母細胞,且能夠被定向誘導(dǎo)分化為軟骨細胞,人類軟骨糖蛋白39對其分化過程具有一定的促進作用。
[Abstract]:Background: studies have shown that human cartilage glycoprotein 39 has a certain relationship with the degeneration and repair of osteoarticular cartilage. Objective: to observe the effect of human chondroglycoprotein 39 on the differentiation of articular cartilage cells induced by human knee joint grandmother chondrocytes. Methods: adult articular cartilage was obtained. Digestion, isolation and culture of articular cartilage grandmother cells; flow cytometry analysis of the amount of cells capable of expressing CD105 and CD166 in the passage cells; isolation and purification of the isolated chondrograndmother cells. The isolated chondrograndmother cells were cultured by monolayer culture. After subculture to the second generation, the chondrograndmother cells were cultured in the cartilage medium containing human chondroglycoprotein 39 and common cartilage induction medium for 14 days, respectively. The expression of type 鈪,

本文編號:1571455

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