培养条件对CD34^+细胞体外诱导分化和扩增红细胞的影响  

Influence of culture conditions on proliferation and differentiation of induced red blood cells from CD34^+ cells in vitro

作　　者：贾延军[1] 刘江[1] 张可莹[1] 单小燕[1] 李伟[1] 何晓梅[1] 王立君[1] 刘娜[1] 王琳[1] 崔爽[1] 倪雷[1] 赵博涛[1] 龚志尹[1] 王东玫[1] 高颂明[1] 张志欣[1] 

机构地区：[1]北京市红十字血液中心,北京100088

出　　处：《中国输血杂志》2010年第2期107-110,共4页Chinese Journal of Blood Transfusion

摘　　要：目的观察培养基以及细胞接种密度对体外诱导CD34+细胞向红细胞增殖和分化的影响。方法将脐血来源的CD34+细胞用无血清培养基或含10%胎牛血清(FCS)的IMDM培养基培养(6孔培养板,3ml/孔),添加的细胞因子为100ng/ml干细胞因子(SCF)、5ng/ml白细胞介素3(IL-3)、3IU/ml促红细胞生成素(EPO),细胞初始接种密度为104/ml或105/ml,并通过间充质干细胞(MSC)的支持培养来诱导其向红系细胞扩增和分化。结果培养23d后,无血清培养基组细胞扩增倍数达到2.54×105倍,其中红系细胞>95%,而IMDM/5%FCS组的最大扩增倍数只有1.84×104倍,其中的红系细胞约占40%。培养8d后初始接种密度为104/ml的CD34+细胞增殖(420±40)倍,而105/ml的CD34+细胞增殖(162±45)倍。结论CD34+细胞在无血清培养基中的扩增倍数以及最终诱导出的红细胞,都明显高于在IMDM/10%FCS培养基中所占比例,104/ml的CD34+细胞接种密度比105/ml更有利于细胞的扩增。Objective To investigate the influence of medium and initial cell concentration on proliferation and differentiation of cultured red blood cells from CD34^＋ cells in vitro.Methods Cord blood derived CD34^＋ cells were cultured in serum-free medium or Iscove＇s modified Dulbecco＇s medium（IMDM）containing 10% fetal calf serum（FCS）supplemented with 100 ng/ml of stem cell factor（SCF）,5 ng/mL of interleukin-3（IL-3）and 3 IU/mL erythropoietin（EPO）and induced to differentiate into erythroid cells by co-cultured with human mesenchymal stem cells（MSC）derived from bone marrow（BM）.Results After 23 days of culture,the cells cultured in serum-free medium were amplified at（2.54×10^5）folds in which erythroid cells accounted for over 95%,compared with（1.84×10^4）folds and about 40% of erythroid cells cultured in IMDM/5%FCS group.By day 8（420 ± 40）folds of proliferation could be accomplished for CD34^＋ cells with the initial concentration of 104/ml,higher than the（162 ± 45）folds for CD34^＋ cells with the initial concentration of 105/ml.Conclusion Selection of suitable medium was critical for the induction of red blood cells in vitro.Initial inoculating density of 104/ml of CD34^＋ is preferred for amplification of erythroid cells to 10^5/ml.

关 键 词：红细胞 体外培养 CD34^+细胞 接种密度 增殖 分化 培养基 

分 类 号：Q813.11[生物学—生物工程] R329.28[医药卫生—人体解剖和组织胚胎学]