Duan Hua-Xin, Mao Ping, Deng Ting-Fen, Luo Chang-Ru, Xu Yan-Li, Zhang Yu-Ping
Department of Hematology, The First Guangzhou Municipal People Hospital, Guangzhou Medical College, Guangzhou 510180, Guangdong Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2008 Aug;16(4):867-70.
The aim of this study was to expand hematopoietic progenitor cells at large scale by magnet stirred culture system. Mononuclear cell from umbilical cord blood were cultured in serum-free medium with stem cell factor, FIT-3 ligand and thrombopoietin. Firstly, the role of magnet on cell growth and colony-forming was studied by static culture on 0, 25 and 50 mT. Then the expansion multiple of cells, colony-forming and expression of surface markers were studied in magnet stirred culture by cell counting, colony-forming assay and flow cytometry. The results indicated that there was no difference in multiple of total cell expansion and numbers of hematopoietic colonies between 0, 25 and 50 mT groups and spinner groups (all p > 0.05). After 7 day cultures, the multiple of total cell expansion in magnet stirred culture was higher than that in static culture (p < 0.01). The numbers of CFU-GM (colony-forming unit-granulocyte/macrophage) and CFU-E (erythroid colony forming unit) in magnet stirred culture were higher than those in static culture, (p < 0.05). The primitive cells (CD34(+), CD34(+)/CD38(-) or CD133(+)) of the expanded cells in magnet stirred culture were less than those in static culture (p < 0.05). However, the CD184(+) or CD62L(+) expanded cells were more than that in static culture (p < 0.05). It is concluded that magnet stirred culture favors the expansion of hematopoietic progenitor cells. The results will be finally confirmed in further in vivo experiments and clinical applications.
本研究的目的是通过磁力搅拌培养系统大规模扩增造血祖细胞。将脐血单个核细胞在含干细胞因子、FIT-3配体和血小板生成素的无血清培养基中培养。首先，通过在0、25和50 mT下进行静态培养研究磁场对细胞生长和集落形成的作用。然后通过细胞计数、集落形成试验和流式细胞术研究磁力搅拌培养中细胞的扩增倍数、集落形成和表面标志物的表达。结果表明，0、25和50 mT组与旋转培养组之间在总细胞扩增倍数和造血集落数量上无差异（所有p>0.05）。培养7天后，磁力搅拌培养中总细胞扩增倍数高于静态培养（p<0.01）。磁力搅拌培养中CFU-GM（粒-巨噬细胞集落形成单位）和CFU-E（红系集落形成单位）的数量高于静态培养（p<0.05）。磁力搅拌培养中扩增细胞的原始细胞（CD34(+)、CD34(+)/CD38(-)或CD133(+)）少于静态培养（p<0.05）。然而，CD184(+)或CD62L(+)扩增细胞多于静态培养（p<0.05）。结论是磁力搅拌培养有利于造血祖细胞的扩增。这些结果最终将在进一步的体内实验和临床应用中得到证实。
