Chi Zhan-You, Xia Quan-Ming, Kang Zi-Zhen, Tan Wen-Song, Dai Gan-Ce
The State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China.
Sheng Wu Gong Cheng Xue Bao. 2003 Sep;19(5):587-92.
Stirred culture offers a number of advantages over static systems as it maintains a stable, homogeneous culture environment and is easy to scale-up. This paper focused on the development and application of stirred tank bioreactor to culture hematopoietic cells. Preliminary study of stirred culture of hematopoietic cells was carried out in cord blood mononuclear cells culture in spinner flask. The results showed that the amplification rates of total cell, CFU-GM and BFU-E, with the exception of CFU-Mk, were greater in spinner flask than T-flask. The number of total cells increased 20 fold after 14 days incubation in spinner flask. The amplification rates of CFU-GM, CFU-Mk and BFU-E reached maximum at 10th day, 10th day and 7th day respectively, and the maximal amplification rates were 9.2-fold, 5.5-fold and 2.4-fold respectively, whereas the rate of CD34+ cells in spinner flask was (6.7 +/- 4.0)-fold at day 10. These results indicated that the stirred culture system is better than the static culture systems for hematopoietic cell proliferation. The biocompatibility of cord blood MNC to different types of materials used in bioreactors was also tested. The results showed that glass, stainless steel 316L and polytetraflouroethylene (PTFE) supported the growth of hematopoietic cells well. A higher cell density was reached in stirred bioreactors with controlled pH and DO than static culture. These findings suggested that the controlled large-scale culture could be used to overcome the clinical shortage of hematopoietic cells.
搅拌培养相对于静态系统具有许多优势，因为它能维持稳定、均匀的培养环境且易于放大规模。本文重点研究了搅拌罐生物反应器在培养造血细胞方面的开发与应用。在转瓶中对脐血单个核细胞进行了造血细胞搅拌培养的初步研究。结果表明，除CFU-Mk外，转瓶中总细胞、CFU-GM和BFU-E的扩增率高于T型瓶。在转瓶中培养14天后，总细胞数增加了20倍。CFU-GM、CFU-Mk和BFU-E的扩增率分别在第10天、第10天和第7天达到最大值，最大扩增率分别为9.2倍、5.5倍和2.4倍，而转瓶中第10天CD34+细胞的扩增率为(6.7±4.0)倍。这些结果表明，搅拌培养系统在造血细胞增殖方面优于静态培养系统。还测试了脐血单个核细胞对生物反应器中不同类型材料的生物相容性。结果表明，玻璃、316L不锈钢和聚四氟乙烯（PTFE）对造血细胞的生长支持良好。与静态培养相比，在pH和溶解氧可控的搅拌生物反应器中可达到更高的细胞密度。这些发现表明，可控的大规模培养可用于克服造血细胞的临床短缺问题。
