郑琛,孙海英,齐念民
ZHENG Chen,SUN Hai-ying,QI Nian-min(School of Pharmacy

• 1:上海交通大学药学院

摘要(Abstract)：

目的探讨大鼠骨髓间充质干细胞(Mesenchymal stem cells,MSCs)培养过程中即时监测细胞浓度的可行性。方法将6～8周龄的SD大鼠处死,分离MSCs,在24孔板上半连续灌注培养和自制的生物反应器上连续灌注培养,通过实时检测培养过程中的葡萄糖消耗率(Glucose consumption rate,GCR)、乳酸累积率(Lactic acid production rate,LPR)及相应的干细胞浓度,并计算葡萄糖的比细胞得率(qGlu)及乳酸累积比细胞得率(qLac),分析其稳定性及细胞浓度与GCR和LPR之间的关系。结果所分离培养的MSCs形态均一,轮廓鲜明,呈典型的短小梭形。在24孔板上半连续灌注培养的qGlu和qLac分别为1.84和1.74 mg/(106个.d);生物反应器连续灌注培养的qGlu和qLac分别为1.66和1.61 mg/(106个.d),二者比细胞得率均为恒值。结论 MSCs培养过程中,GCR、LPR与细胞浓度具有良好的线性关系,通过对葡萄糖浓度和乳酸浓度的测定,可以实时监测MSCs的浓度。
Objective To investigate the feasibility of real-time monitoring the culture concentration of rat mesenchymal stem cells(MSCs).Methods MSCs were isolated from SD rats aged 6 ～ 8 weeks,and subjected to semi-continuous perfusion culture on 24-well microtiter plate and continuous perfusion culture in bioreactor prepared by the authors respectively,and analyzed for stability as well as the relationship of cell concentration to glucose consumption rate(GCR)and lactic acid production rate(LPR)by real-time monitoring the GCR,LPR and the concentration of corresponding MSCs and calculating the specific cell yields of glucose(qGlu)and lactic acid(qLac).Results The cultured MSCs showed homogeneous morphology and clear border,and in typically short shuttlelike form.The qGlu and qLac of MSCs in semi-continuous perfusion culture on 24-well microtiter plate were 1.84 and 1.74 mg /(106 cells.d),while those in continuous perfusion culture in bioreactor were 1.66 and 1.61 mg /(106 cells.d),respectively,all of which were constant values.Conclusion During culture of MSCs,both GCR and LPR showed good linear relationship to cell concentration.The real-time monitoring of concentration of MSCs may be achieved by determination of glucose and lactic acid concentrations.

关键词(KeyWords)： 间质干细胞;细胞浓度;葡萄糖消耗率;乳酸累积率;生物反应器;比细胞得率
Mesenchymal stem cells(MSCs);Cell concentration;Glucose consumption rate(GCR);Lactic acid production rate(LPR);Bioreactor;Specific cell yield

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 郑琛,孙海英,齐念民
ZHENG Chen,SUN Hai-ying,QI Nian-min(School of Pharmacy

DOI: 10.13200/j.cjb.2012.01.116.zhengch.005

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