摘要
目的为骨组织工程的临床应用提供一种新的人骨髓基质干细胞(BMSCs)培养方法,以满足细胞培养过程中对各类细胞因子的需求,同时尽量减少细胞培养过程中动物源性抗原物质的引入。方法采用10%富血小板血浆(PRP)替代动物血清配比高糖DMEM培养基,体外诱导培养(50μg/mL抗坏血酸、10-8mol/L地塞米松、10-3mol/Lβ-甘油磷酸钠)人BMSCs,快速扩增后,倒置相差显微镜、扫描电镜观察各组细胞形态及细胞增殖情况。ALP染色与钙结节染色等方法对细胞进行生物学特性检测。结果人BMSCs24h开始贴壁,7d左右细胞融合。诱导培养后细胞能较快地扩增;ALP染色与钙结节染色结果显示细胞具有良好的成骨细胞生物学特性。结论以自体PRP替代动物血清体外诱导培养人BMSCs是一种良好的培养方法,所培养的细胞数量及其生物学特性能快速达到临床应用的需求。
Objective To discuss a new method of BMSCs culture which can reduce the lead-in of extraneous antigenic substances in the process of cell culture, and can thus better meet the demand on the quantity and biological characteristics of BMSCs in the clinical use of tissue engineering bone. Methods BMSCs from a human adult in vitro were induced by using autologous platelet-rich plasma instead of animal serum in high-carbohydrate DMEM （Dulbecco＇s minimum essential medium） . After the cultured BMSCs were amplified rapidly by Rotary Cell Culture System, their growth status was observed and their biological characteristics were detected. Results The cells were in good growth status and presented fine biological characteristics. Their growth speed in Rotary Cell Culture System was observed to be evidently faster than that by conventional methods. Conclusions Rotary Cell Culture System by autologous platelet-rich plasma instead of animal serum is a good culture method to induce human BMSCs in vitro. The quantity and biological characteristics of BMSCs cultured in this way can meet the demand in clinical use.
出处
《中华创伤骨科杂志》
CAS
CSCD
2006年第1期56-58,共3页
Chinese Journal of Orthopaedic Trauma
基金
国家高科技研究发展计划(863)重点项目(2003AA205001)
广东省自然科学基金项目(04300723)
关键词
骨
组织工程
骨髓基质干细胞
富血小板血浆
Bone
Tissue engineering
Bone marrow stromal cells （BMSCs）
Platelet-rich plasma （PRP）
