Sorting,identification and enrichment of side population cells in acute monocytic leukemia THP-1 Cells
10.3760/cma.j.issn.2095-428X.2013.09.018
- VernacularTitle:急性单核细胞白血病细胞系THP-1侧群细胞的分选、鉴定与富集
- Author:
Ying-Chao WANG
1
;
Li-Na MA
;
Chu-Yun YIN
;
Lei FENG
;
Yong-Wei WEI
;
Guang-Yao SHENG
Author Information
1. 450052,郑州大学第一附属医院儿科
- Keywords:
THP-1 cell line;
Leukemia stem cells;
Side population cells;
Ara-C
- From:
Chinese Journal of Applied Clinical Pediatrics
2013;28(9):699-703
- CountryChina
- Language:Chinese
-
Abstract:
Objective To assay and determine whether the human acute monocytic leukemia cell line THP-1 contains side populations (SP) cells,and to increase the proportion of SP cells using cytarabine (Ara-C).Methods Fluorescent microscope and flow cytometry (FCM) were employed for detecting the percentage of SP cells in THP-1 cells.Then,SP and non-SP (NSP) subpopulations were collected and identified.Finally,THP-1 cells were incubated with different concentrations of Ara-C for 24 hours and detected the proportion of SP cells,respectively.Results The results demonstrated that the percentage of SP cells was (1.81 ± 0.99) ％ in THP-1 cells.A majority of the SP cells remained in the G0/G1 phase,and the expressions of CD34 + and CD34 + CD38-and the proliferative ability of the SP cells were higher than those of NSP cells (P ＜ 0.05).The mRNA expression of multidrug resistance genes (ABCG2,ABCB1),apoptosis regulation genes (Bcl-2) and the Bcl-2/Bax ratio of SP cells were higher than those of NSP cells.SP cells have been shown to be more tumorigenic than NSP cells.After co-culture with Ara-C,the proportion of SP cells increased significantly and presented in a concentration-dependent manor.Conclusions All of these findings suggest that the THP-1 cell line contains SP cells and the SP cells possess some intrinsic stem cell properties.The proportion of SP cells can be increased when co-cultured with Ara C,and this technique is a useful and important application for the study of LSCs.
