大鼠骨髓间充质干细胞培养浓度的即时监测Real-time monitoring of culture concentration of rat mesenchymal stem cells
郑琛;孙海英;齐念民;
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。
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
基金项目(Foundation):
作者(Author): 郑琛;孙海英;齐念民;
Email:
DOI: 10.13200/j.cjb.2012.01.116.zhengch.005
参考文献(References):
- [1]Ringe J,Kaps C,Schmitt B,et al.Porcine mesenchymal stemcells.Induction of distinct mesenchymal cell lineages[J].CellTissue Res,2002,307(3):321-327.
- [2]姚春丽,夏建新,陈玉丙.骨髓间充质干细胞对豚鼠皮肤光老化的修复作用[J].中国生物制品学杂志,2008,21(4):315-317.
- [3]李秀东,陈玉丙,宋祥福,等.小鼠骨髓间充质干细胞对异基因肝移植模型大鼠移植区域组织损伤的修复作用[J].中国生物制品学杂志,2008,21(3):221-223.
- [4]Van Damme A,Vanden Driessche T,Collen D,et al.Bone mar-row stromal cells as targets for gene therapy[J].Curr Gene Ther,2002,2(2):195-209.
- [5]Deans RJ,Moseley AB.Mesenchymal stem cells:biology and po-tential clinical uses[J].Exp Hematol,2000,28(8):875-884.
- [6]Pelletier F,Fonteix C,Da Silva AL,et al.Software sensors forthe monitoring of perfusion cultures:Evaluation of the hybridomadensity and the medium composition from glucose concentrationmeasurements[J].Cytotechnology,1994,15(1-3):291-299.
- [7]Cannizzaro C,Gügerli R,Marison I,et al.On-line biomass moni-toring of CHO perfusion culture with scanning dielectric spec-troscopy[J].Biotechnol Bioeng,2003,84(5):597-610.
- [8]Dowd JE,Jubb A,Kwok KE,et al.Optimization and control ofperfusion cultures using a viable cell probe and cell specific perfu-sion rates[J].Cytotechnology,2003,42(1):35-45.
- [9]Ducommun P,Kadouri A,Von Stockar U,et al.On-line determi-nation of animal cell concentration in two industrial high densityculture processes by dielectric spectroscopy[J].Biotechnol Bio-eng,2002,77(3):316-323.
- [10]Kilburn DG,Fritzpatrick P,Blake-Coleman BC,et al.On-linemonitoring of cell mass in mammalian cell cultures by acousticdensitometry[J].Biotechnol Bioeng,1989,33(11):1379-1384.
- [11]欧阳杰,武彦文,王晓东,等.肉苁蓉细胞悬浮培养动力学研究[J].北京林业大学学报,2007,29(5):132-136.
- [12]Ozturk SS,Thrift JC,Blackie JD,et al.Real-time monitoringand control of glucose and lactate concentrations in a mammaliancell perfusion reactor[J].Biotechml Bioeng,1997,53(4):372-378.
- [13]Cruz HJ,Ferreira AS,Freitas CM,et al.Metabolic response todifferent glucose and glutamine levels in baby hamster kidney cell culture[J].Appl Microbiol Biotechnol,1999,51(5):579-585.
- [14]Hu WS,Oberg MG.Monitoring and control of animal cell biore-actors:biochemical engineering considerations[J].BioprocessTechnol,1990,10(3):451-481.
- [15]Zielke HR,Ozand PT,Tildon JT,et al.Reciprocal regulation ofglucose and glutamine utilization by cultured human diploid fi-broblasts[J].J Cell Physiol,1978,95(1):41-48.