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该文采用旋转培养方式研究微重力培养条件对造血干祖细胞向中性粒细胞分化的效率和功能的影响。脐血CD34+细胞用扩增体系[Stem Span SFEM培养基、干细胞因子(stem cell factor,SCF)、Fms相关酪氨酸激酶3受体(Fms-related tyrosine kinase 3 ligand,Flt3)、血小板生成素(thrombopoietin,TPO)、白细胞介素-3(interleukin-3,IL-3)]静置培养7 d后,再分成静置培养(static culture,SC)组和旋转培养(rotate cell culture system,RCCS)组,并换为分化培养体系[Stem Span SFEM培养基、IL-6、粒细胞集落刺激因子(granulocyte colony stimulating factor,G-CSF)、SCF、Flt3、IL-3]再培养9 d。培养过程中用流式细胞术检测CD34、CD38、CD11b、CD16b、CD66b比例,第16 d收集细胞进行中性粒细胞功能检测。结果表明,RCCS组和SC组均能形成形态正常和功能成熟的中性粒细胞,RCCS组CD16b~+的中性粒细胞比例显著增多(29.82%±2.48%vs 15.01%±0.62%,P<0.01),且细胞产生活性氧簇(reactive oxygen species,ROS)的能力显著增强(P<0.05),具有趋化运动能力的细胞增多(50.430%±3.275%vs 34.000%±0.700%,P<0.05)、运动速度增快(8.100±0.404μm/min vs 5.850±0.050μm/min,P<0.05)。该文结果提示,微重力培养条件能促进造血干祖细胞分化为形态正常和功能成熟的中性粒细胞。
In this paper, the effects of microgravity culture on the efficiency and function of hematopoietic stem / progenitor cells to neutrophil differentiation were studied by rotating culture. Umbilical cord blood CD34 + cells were treated with an amplification system [Stem Span SFEM medium, stem cell factor (SCF), Fms-related tyrosine kinase 3 ligand (Flt3), thrombopoietin Thrombopoietin (TPO) and interleukin-3 (IL-3)] were cultured in static condition for 7 days, then divided into static culture (SC) group and rotate cell culture system (RCCS) The cells were re-cultured for 9 days in Stem Span SFEM, IL-6, G-CSF, SCF, Flt3 and IL-3. During culture, the percentage of CD34, CD38, CD11b, CD16b and CD66b was detected by flow cytometry, and the neutrophil function was detected on the 16th day. The results showed that neutrophils with normal morphology and mature function could be formed in both RCCS and SC groups. The proportion of CD16b + neutrophils in RCCS group was significantly increased (29.82% ± 2.48% vs 15.01% ± 0.62%, P < 0.01). The ability of cells to produce reactive oxygen species (ROS) was significantly enhanced (P <0.05) and the number of chemotactic cells increased (50.430% ± 3.275% vs 34.000% ± 0.700%, P <0.05 ), The speed of motion increased (8.100 ± 0.404μm / min vs 5.850 ± 0.050μm / min, P <0.05). The results suggest that micro-gravity culture conditions can promote the differentiation of hematopoietic stem and progenitor cells into normal morphology and mature neutrophils.