Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (10): 1821-1824.doi: 10.3969/j.issn.1673-8225.2010.10.023

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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

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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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