SCF、FL及TPO在低氧条件下扩增脐血单个核细胞  

Effect of stem cell factor （SCF）, Flt3 Ligand （FL） and thrombopoietin （TPO） on ex vivo expansion of mononuclear cells derived from umbilical cord blood under hypoxic conditions

作　　者：许力[1] 莫文健[1] 毛平[1] 

机构地区：[1]广西壮族自治区人民医院血液科,南宁530021

出　　处：《中华器官移植杂志》2015年第7期429-432,共4页Chinese Journal of Organ Transplantation

摘　　要：目的 探讨低氧条件下干细胞生长因子（SCF）、FLT3配基（FL）及促巨核细胞生成因子（TPO）3种细胞因子组合对人脐血单个核细胞体外扩增的效果.方法 采集新鲜人脐血,分离单个核细胞,根据细胞培养时是否加入细胞因子组合及是否低氧条件（体积分数3％）,分4组培养观察,A组为常氧条件下不含细胞因子,B组为低氧条件下不含细胞因子,C组为常氧条件下含SCF、FL和TPO,D组为低氧条件下含SCF、FL和TPO.在培养0d和7d时检测各组培养体系中的有核细胞数（TNC）、CD34＋细胞数、CD34＋CXCR4＋细胞数、CD34＋ CD49d＋细胞数、CD34＋ CD62L＋细胞数及集落形成单位（CFU）数.结果 培养7d时,B组、C组和D组扩增的TNC数、CD34＋细胞数及CFU数等扩增倍数均显著高于A组,与A组比较,差异均有统计学意义（P＜0.05）,而B组与C组间均无明显差异（P＞0.05）,D组的扩增效果均显著优于B组和C组（P＜0.05）.培养7d时,A组CD34＋ CxCR4＋细胞、CD34＋ CD62L＋细胞及CD34＋ CD49d＋细胞数等扩增倍数均较培养0d时下降.与A组相比,B组、C组和D组CD34＋ CxCR4＋细胞、CD34＋ CD62L＋细胞及CD34＋ CD49d＋细胞等扩增倍数均显著高于A组,差异均有统计学意义（P＜0.05）而B组与C组间均无明显差异（P＞0.05）,D组的扩增效果均显著优于B组和C组（P＜0.05）.结论 低氧可促进SCF、FL和TPO组合扩增脐血单个核细胞,并能促进和归巢有关的CD49d、CD62L及CXCR4的表达.Objective To explore the effect of stem cell factor （SCF）, Flt3 Ligand （FL） and Thrombopoietin （TPO） on the expansion of the mononuclear cells derived from umbilical cord blood ex vivo under hypoxic conditions.Method Human fresh CB mononuclear cells were isolated and cultured with different combinations of cytokine for 7 days under normoxic or hypoxic conditions （3 % O2）.At day 0 and 7 of culture, total nucleated cells （TNC）, CD34＋ , CD34＋ CXCR4＋ , CD34＋ CD49d＋ and CD34＋ CD62L＋ cells were assayed by flow cytometry, and colony forming unit （CFU） was determined.The experiment was divided into four groups： group A （cells were cultured in single mononuclear cells under normoxic conditions）;group B （cells were cultured in single mononuclear cells under hypoxic conditions）;group C （cells were cultured with SCF, FL and TPO under normoxic conditions）;group D （cells were cultured with SCF, FL and TPO under hypoxic conditions）.Result At 7th day of culture, TNC, CD34＋ cells and CFU expansion were significantly increased in groups B, C and D, and the expansion effect in group D was superior to that in groups B and C, but there was no significant difference between groups B and C （P〉0.05）.At 7th day of culture, the expression levels of CD49d, CD62L and CXCR4 on expanded CD34 ＋ cells were significantly reduced in group A as compared with those at day 0, and those in groups B, C and D were significantly increased as compared with those in group A, those in group D were significantly higher than in group D than in groups B and C （P〈0.05）,but there was no significant difference between groups B and C （P〉 0.05）.Conclusion Hypoxic conditions could enhance the expansion and expression of CD49d, CD62L and CXCR4 of mononuclear cells derived from umbilical cord blood with SCF, FL and TPO.

关 键 词：脐血 单个核细胞 体外扩增 低氧 干细胞生长因子 FLT3配基 促巨核细胞生成因子 

分 类 号：R457.7[医药卫生—治疗学]