本文選題：骨髓間充質(zhì)干細(xì)胞 + 細(xì)胞培養(yǎng)��； 參考：《中南大學(xué)》2008年碩士論文

【摘要】： 目的建立一種體外無血清培養(yǎng)擴(kuò)增人骨髓間充質(zhì)干細(xì)胞的方法,為骨組織工程研究提供種子細(xì)胞。 方法從5例手術(shù)病人骨髓中分離制備單個核細(xì)胞懸液,分別接種于有血清培養(yǎng)基及自制的無血清培養(yǎng)基中,比較兩種培養(yǎng)條件下細(xì)胞生長情況,流式細(xì)胞儀分析兩種培養(yǎng)條件下間充質(zhì)干細(xì)胞表面CD分子表達(dá)情況及細(xì)胞周期。 結(jié)果有血清培養(yǎng)2～3天后細(xì)胞開始出現(xiàn)貼壁生長,1周左右有細(xì)胞叢生長,12～14天細(xì)胞融合成片,呈典型的旋渦狀排列。無血清培養(yǎng)4～5天后細(xì)胞開始出現(xiàn)貼壁生長,8～10天左右有細(xì)胞叢生長,16～20天細(xì)胞融合成片。單層融合的MSCs消化傳代培養(yǎng),有血清培養(yǎng)1周左右達(dá)融合,無血清培養(yǎng)9～10天左右達(dá)融合,可繼續(xù)傳代擴(kuò)增。流式細(xì)胞儀檢測顯示,兩種培養(yǎng)條件下擴(kuò)增后的人骨髓MSCs均不表達(dá)CD14,CD34和CD45,而強(qiáng)表達(dá)CD29、CD54、CD105;無血清培養(yǎng)組G_0/G_1期細(xì)胞比例明顯高于有血清培養(yǎng)組(P＜0.05)。 結(jié)論 1.利用自行配制的無血清培養(yǎng)基能較好地培養(yǎng)、擴(kuò)增人骨髓MSCs。 2.無血清培養(yǎng)組G_0/G_1期細(xì)胞比例明顯高于有血清培養(yǎng)組,說明與有血清培養(yǎng)相比,無血清培養(yǎng)的細(xì)胞其干細(xì)胞特性保存更好。
[Abstract]:Objective to establish a method for amplification of human bone marrow mesenchymal stem cells (BMSCs) by serum-free culture in vitro to provide seed cells for bone tissue engineering. Methods Mononuclear cell suspensions were isolated from bone marrow of 5 patients and inoculated in serum-free medium and serum-free medium respectively. The expression and cell cycle of CD molecules on mesenchymal stem cells were analyzed by flow cytometry. Results after 2 days of serum-culture, the cells began to grow with adherent cells for 1 week or so. After 1214 days of cell growth, the cells were fused into pieces, and the cells were arranged in a typical swirl shape. After 5 days of serum-free culture, adherent growth began to occur in the cells. In monolayer fusion MSCs digestion and passage culture, serum-free culture reached fusion about 1 week, serum-free culture reached fusion about 10 days, and can continue to be subcultured and amplified. The results of flow cytometry showed that human bone marrow MSCs did not express CD14 CD34 and CD45, but strongly expressed CD29, CD54 and CD105, and the proportion of G_0/G_1 phase cells in serum-free culture group was significantly higher than that in serum-free culture group (P < 0.05). Conclusion 1. Human bone marrow MSCs could be amplified by using self-made serum-free medium. 2. The proportion of G_0/G_1 phase cells in serum-free culture group was significantly higher than that in serum-free culture group, which indicated that the stem cell characteristic of serum-free culture group was better than that of serum-free culture group.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R329

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