CAJ | 학술논문

目的优化脐血单核细胞( UCB-MNCs)的培养方法.方法采用正交实验方法筛选影响UCB-MNCs培养的各因素,优化培养条件.设立优化组、对照组(UCB-MNCs以1.0×106/L接种于含20 μg/L碱性成纤维细胞生长因子(bFGF)、体积分数为0.10胎牛血清、低糖DMEM培养基)培养.其后检测各组培养细胞增殖及形态学变化；流式细胞仪对各组行免疫表型分析.结果筛选后优化培养条件为:低糖DMEM培养基中含20 mg/L bFGF、50 μg/L干细胞因子(SCF)、10 μg/L重组人粒细胞生长因子(G-CSF)、培养瓶用层粘连蛋白(LN)包被.原代培养的细胞密度优化组细胞( 108.15±6.90)增殖优于对照组(51.48±12.27),差异有统计学意义(P＜0.01).P3代UCB-MNCs中MSCs含量优化组(94.87％)优于对照组(75.35％),差异有统计学意义(P＜0.05).结论采用正交实验的方法可以筛选出脐血单核细胞的优化培养条件.
목적 우화제혈단핵세포( UCB-MNCs)적배양방법.방법 채용정교실험방법사선영향UCB-MNCs배양적각인소,우화배양조건.설립우화조、대조조(UCB-MNCs이1.0×106/L접충우함20 μg/L감성성섬유세포생장인자(bFGF)、체적분수위0.10태우혈청、저당DMEM배양기)배양.기후검측각조배양세포증식급형태학변화；류식세포의대각조행면역표형분석.결과 사선후우화배양조건위:저당DMEM배양기중함20 mg/L bFGF、50 μg/L간세포인자(SCF)、10 μg/L중조인립세포생장인자(G-CSF)、배양병용층점련단백(LN)포피.원대배양적세포밀도우화조세포( 108.15±6.90)증식우우대조조(51.48±12.27),차이유통계학의의(P＜0.01).P3대UCB-MNCs중MSCs함량우화조(94.87％)우우대조조(75.35％),차이유통계학의의(P＜0.05).결론 채용정교실험적방법가이사선출제혈단핵세포적우화배양조건.
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.