中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (10): 1821-1824.doi: 10.3969/j.issn.1673-8225.2010.10.023

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

血液肿瘤细胞株中侧群细胞的检测和分选

范瑞华1，李惠民2，李晓进2，喻镁佳2，金从国3   

  1. 1淮安市第一人民医院肿瘤科，江苏省淮安市  223300；
    2昆明医学院第一附属医院血液科，云南省昆明市  650032；
    3昆明医学院第三附属医院肿瘤研究所，云南省昆明市  650031
  • 出版日期:2010-03-05 发布日期:2010-03-05
  • 通讯作者: 李惠民，教授，昆明医学院第一附属医院血液科，云南省昆明市 650032 lihuimin@ medmail.com.cn
  • 作者简介:范瑞华，女，1977年生，山西省左权县人，汉族，2009年昆明医学院毕业，硕士，医师，主要从事血液肿瘤的诊断和治疗。 fanruihuabaobao@163.com

Detection and separation of side population cells in haematological tumor cell lines

Fan Rui-hua1, Li Hui-min2, Li Xiao-jin2, Yu Mei-jia2, Jin Cong-guo3   

  1. 1Department of Oncology, First People’s Hospital of Huaian, Huaian  223300, Jiangsu Province, China;
    2Department of Hematology, the First Affiliated Hospital of Kunming Medical College, Kunming  650021, Yunnan Province, China;
    3Institute of Oncology, Third Affiliated Hospital of Kunming Medical College, Kunming  650031, Yunnan Province, China
  • Online:2010-03-05 Published:2010-03-05
  • Contact: Li Hui-min, Professor, Department of Hematology, First Affiliated Hospital of Kunming Medical College, Kunming 650021, Yunnan Province, China lihuimin@medmail. com.cn
  • About author:Fan Rui-hua, Master, Physician, Department of Oncology, First People’s Hospital of Huaian, Huaian 223300, Jiangsu Province, China fanruihuabaobao@ 163.com

PDF

461

被引次数

17

摘要：

背景：侧群细胞广泛分布在多种成体组织，胚胎和某些肿瘤细胞系中，在不同的成体组织中含量不尽相同。血液肿瘤是危害人类的主要疾病之一，在血液肿瘤进行侧群细胞分布和方法研究对认识侧群细胞有重要的意义。

目的：鉴定血液肿瘤的4个细胞株中是否存在侧群细胞，并掌握稳定的检测和分选侧群细胞的方法。

方法：利用干细胞高效能将Hoechst33342荧光染料泵出细胞的特性，采用流式细胞仪检测在NB4、Raji、K562/ADM和K562细胞株中是否存在侧群细胞；对分选后K562两亚群细胞进行侧群细胞检测，以检测分选细胞的纯度。

结果与结论：经Hoechst33342染色，流式细胞仪分析结果显示在4个血液肿瘤细胞株NB4、Raji、K562/ADM和K562细胞株中均存在侧群细胞，这群细胞比例均少，分别为0.8%、2.7%、1.3%和2.7%，能被Verapamil完全抑制。分选K562细胞侧群和非侧群细胞再次检测发现分选细胞纯度高。

关键词: 白血病干细胞, 侧群细胞, 流式细胞术, 肿瘤干细胞, 细胞株

Abstract:

BACKGROUND: Side population (SP) cells, with varied contents, are widely distributed in adult tissues, embryos, and certain tumor cells. Haematological tumor is one of the main pathological conditions, which endangers human life. Thus, it is important to recognize SP cells in haematological tumor cell lines.

OBJECTIVE: To identify whether hematologic tumor cell lines contain SP cells, and to explore a stable detection and separation methods.

METHODS: Cells with the characteristics of stem cells being capable of fluorescent dye Hoechst33342 were isolated by flow cytometry; whether there were SP cells in logarithmic growth period of NB4, Raji, K562/ADM and K562 or not were detected. After sorting K562 subpopulations, the purity of sorted cells was detected. 

RESULTS AND CONCLUSION:After Hoechst33342 staining, flow cytometry results showed that the SP cells appeared in the haematological tumor NB4, Raji, K562/ADM and K562 cell lines, which accounted for 0.8%, 2.7%, 1.3% and 2.7%, respectively. These cells could be blocked by Verapamil. The purity was greater after a second detection of SP and Non SP cells in K562 cells.

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