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醫(yī)學(xué)免費(fèi)論文:骨髓基質(zhì)細(xì)胞體外三維培養(yǎng)的研究
【摘要】 目的：建立骨髓基質(zhì)細(xì)胞(BMSC)體外三維培養(yǎng)的方法，并觀察其細(xì)胞生長(zhǎng)形態(tài)及生物學(xué)特征。方法：制備鼠尾膠原，對(duì)BMSC進(jìn)行三維培養(yǎng)，共聚焦顯微鏡觀察細(xì)胞形態(tài)，MTT、ALP鈣鈷、蘇丹Ⅲ染色觀察BMSC增殖及分化。結(jié)果：三維培養(yǎng)的BMSC早期呈現(xiàn)集落生長(zhǎng)特點(diǎn)，后逐漸向不同的形態(tài)轉(zhuǎn)變，與二維培養(yǎng)相比，細(xì)胞增殖峰值提前，且向成骨及脂肪細(xì)胞分化能力增強(qiáng)。結(jié)論：BMSC在體外三維培養(yǎng)模式下生長(zhǎng)良好，能夠繼續(xù)保持其自我更新能力及分化潛能。
【關(guān)鍵詞】 骨髓基質(zhì)細(xì)胞;三維培養(yǎng)
Three-dimensional Cultivation of Bone Marrow Stromal Cells in vitroWU Yan，YANG Jian，ZHANG Hong-kao，LIU Ji-jun,LI Dong-sheng (Department of Cardiology;Department of Orthopedics，Renmin Hospital,Yunyang Medical College,Shiyan,Hubei 442000,China)Abstract:Objective To establish a method for three-dimensional cultivation of bone marrow stromal cells(BMSCs) in vitro, and to observe its growth morphology and biological characteristics.Methods BMSC were cultured with prepared collagen in a manner of three-dimensional cultivation.The morphology of BMSCs was observed under confocal microscopy,the proliferation and differentiation of BMSCs were detected with MTT,Ca-co staining and sudan-Ⅲ staining.Results BMSCs grew as colony-like at early stage,and then differentiated to different forms.Compared with two-dimensional cultivation,the peak of proliferation was in advance,and the capacity of differentiation to osteoblast and adipocyte was strengthened.Conclusion Three-dimensional cultivation of BMSCs could maintain their capacity of self-renewal and differentiation potential,and cells could grow very well醫(yī).學(xué).全.在.線www.gydjdsj.org.cn.
Key words:Bone marrow stromal cell;Three-dimensional cultivation
骨髓基質(zhì)細(xì)胞因其在二維培養(yǎng)中具有較強(qiáng)的自我更新能力及多分化潛能，而被認(rèn)為是基因治療中合適的種子細(xì)胞。但是細(xì)胞二維培養(yǎng)與體內(nèi)環(huán)境有較大差別，沒(méi)有了基質(zhì)的支持，細(xì)胞貼壁生長(zhǎng)，可能會(huì)失去原有的形態(tài)特征及生長(zhǎng)分化能力。三維培養(yǎng)系統(tǒng)為細(xì)胞提供了立體的生長(zhǎng)空間，能更好的模擬細(xì)胞在體內(nèi)的生理狀態(tài)。本實(shí)驗(yàn)在體外模擬體內(nèi)環(huán)境,構(gòu)建BMSCs三維培養(yǎng)體系，使細(xì)胞所處的環(huán)境盡可能接近體內(nèi),進(jìn)一步研究BMSCs的生物學(xué)性能。
