Abstract
Objective To optimize the culture method of umbilical cord blood-mononuclear cells (UCB-MNCs). Methods First,crosscut empirical method was employed to sieve culture factors of UCB-MNCs,optimize the culture condition,Set up optimize group and control group [ UCB-MNCs was plated in 1.0 × 106/L Lo CHO DMEM supplemented with 20 μg/L basic fibroblast growth factor ( bFGF),10％ FBS)].Then generation and morphology was detected.Immunophenotype was analysesed by FCM.Results The sieved optimized cultivate condition:culture flask was coating by laminin (LN),UCB-MNCs was plated in it ( 1.0 × 106/L) containing Lo CHO DMEM supplemented with 10 μg/L granulocyte colony stimulating factor (G-CSF),50 μg/L stem cell factor (SCF),20 μg/L bFGF,20 ml/L B27.Optimize group ( 108.15 ±6.90) Compared to the control group (51.48 ± 12.27),cell proliferation was more significant (P ＜ 0.01 ),P3 UCB-MNCs content more MSCs in optimized group.Conclusion Optimized cultivate condition of UCB-MNCs could be sieved by orthogonal experiment. Key words: Umbilical cord blood-mononuclear cells; Culture
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