脐血单个核细胞的搅拌悬浮培养

Stirred Culture of Umbilical Cord Blood Mononuclear Cells

研究了悬浮培养中接种密度、换液和优化培养中所用细胞因子组合对造血细胞扩增的影响。结果表明,较高的接种密度(1.0×106cells/mL)比低接种密度(0.5×106cells/mL)更有利于总细胞、粒巨噬集落形成单位(CFU-GM)和巨核细胞集落形成单位(CFU-Mk)的扩增。换液能及时补充葡萄糖和移出培养中产生的乳酸,从而不对细胞生长造成影响,而且可以成功地控制pH不超出造血细胞正常生长所需pH范围。在SCF、IL-3和IL-6组合基础上添加Flt-3配体(FL)和促血小板生成素(TPO)能比原来的SCF、IL-3和IL-6组合更能促进总细胞、CFU-GM和CFU-Mk的扩增。

Stirred culture is more promising than static culture in the large-scale ex vivo culture of hematopoietic cells. The effects of inoculum cell density, medium exchanging, and cytokine combination on expansion of hematopoietic cells in spinner flask culture were studied in this paper. Results indicated that high density of inoculum(1.0×10~6 cells/mL)resulted in more expansion of total cell, CFU-GM and CFU-Mk than that with low inoculum (0.5×10~6 cells/mL). By exchanging medium, the concentration of glucose and lactate and pH can be maintained in the normal range that is suitable to growth of hematopoietic cells. The cytokine combination of FL+TPO+SCF+IL-3+IL-6 could get more expansion of total cell, CFU-GM and CFU-Mk than the control that only SCF+IL-3+IL-6 were added.