中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (45): 8460-8463.doi: 10.3969/j.issn.1673-8225.2010.45.023

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

影响外周血CD34+细胞纯化的相关因素

白  雯，白  槟，王黎明，周建军，张  聪，刘  涛   

  1. 解放军第三二三医院细胞治疗中心，陕西省西安市   710054
  • 出版日期:2010-11-05 发布日期:2010-11-05
  • 通讯作者: 王黎明，硕士，主任医师，解放军第三二三医院细胞治疗中心，陕西省西安市 710054 wanglm@fmmu.edu.cn
  • 作者简介:白雯，女，1965年生，陕西省西安市人，汉族，2000年解放军第三军医大学检验系毕业，副主任技师，主要从事临床检验研究。 bai_wen_323@126.com

Related factors affecting peripheral blood CD34+ cell purification

Bai Wen, Bai Bin, Wang Li-ming, Zhou Jian-jun, Zhang Cong, Liu Tao　   

  1. Cell Therapy Center, the 323 Hospital of Chinese PLA, Xi’an  710054, Shaanxi Province, China 　
  • Online:2010-11-05 Published:2010-11-05
  • Contact: Wang Li-ming, Master, Chief physician, Cell Therapy Center, the 323 Hospital of Chinese PLA, Xi’an 710054, Shaanxi Province, China wanglm@fmmu.edu.cn
  • About author:Bai Wen, Associate chief technician, Cell Therapy Center, the 323 Hospital of Chinese PLA, Xi’an 710054, Shaanxi Province, China bai_wen_323@126.com

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摘要：

背景：造血干细胞具有良好的自我复制和更新的能力，CD34+细胞具备有造血干细胞的标志。
目的：分析影响外周血CD34+细胞纯化的因素。
方法：90例患者，经粒细胞集落刺激因子5 μg/(kg•d)动员1~3 d后，应用COBE血细胞分离机采集外周血单个核细胞液80~100 mL，经Clini MACS免疫磁珠分选技术纯化CD34+细胞。
结果与结论：90例CD34+细胞平均数为(1.73±1.15)×107，经流式细胞仪分析，CD34+细胞阳性率大于80%。COBE血细胞分离机单次收集的循环血量在980~1 100 mL时，利于CD34+细胞收集(P=0.005)；动员后白细胞浓度在(16~21)×109 L-1时，利于CD34+细胞收集(P < 0.05)；中间细胞和淋巴细胞总比例超11%时，利于CD34+细胞收集(P < 0.05)；单个核细胞液血小板小于2 100×109 L-1时，利于CD34+细胞的收集(P < 0.05)；年龄小于16岁，CD34+细胞数高(P=0.003)；CD34+细胞抗体的温度、磁性标记及细胞处理时离心力的大小，均有影响。结果提示，经Clini MACS免疫磁珠细胞分选技术纯化的CD34+细胞能满足临床需要，实验稳定性好，重复性好；注重相关因素的影响，可提高纯化的CD34+细胞数量。

关键词: 外周血, CD34+细胞, 纯化, 免疫磁珠分选技术

Abstract:

BACKGROUND: Hemopoietic stem cells have the potential of self replication and renewal. CD34+ cells have the marker of hemopoietic stem cells.
OBJECTIVE: To analyze the related factors that affect the peripheral blood CD34+ cell purification.
METHODS: From 90 cases of patients, the peripheral blood were collected, after human granulocyte colony stimulating factor (G-CSF) 5 μg/kg•d mobilizing for 1-3 days, using COBE machine mononuclear cells fluid 80-100 mL was collcected, the CD34+  stem cells were purified by Clini MACS.
RESULTS AND CONCLUSION: The mean value of CD34+ cells was (1.73±1.15)×107 in 90 cases. Using flow cytometry, the positive rate of CD34+ cells was more than 80%. When the circulation blood liquid singly collected by COBE machine was 980- 1 100 mL, it was useful for CD34+ cells collection (P = 0.005). After mobilizing, when the white blood cell concentration was (16-21)×109/L, it was useful for CD34+ cells collection (P < 0.05);  when the general ratio of middle cells to lymphocytes was more than 11%, it was useful for CD34+ cell collection (P < 0.05).  When the concentration of blood platelet in mononuclear cells was less than 2 100×109/L, it was useful for CD34+ cell collection (P < 0.05); when the patient’s age was younger than 16 years, the number of collected CD34+ cells was high (P = 0.003). Moreover, the temperature of CD34+ cells, the magnetism marker, the eccentricity of treating cells, all could affect the CD34+ cells collection. Results indicate that the CD34+ cells purified by Clini MACS machine could fit the clinical need and keep good experiment stability and repeatability. To improve the CD34+ cells collection, the related factors which affect the peripheral blood CD34+ purification should be carefully considered.

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