Sorting, identification and enrichment of side population cells in THP-1 acute monocytic leukemia cells

Wang,Yingchao; Yin,Chuyun; Feng,Lei; Ma,Lina; Wei,Yongwei; Sheng,Guangyao

doi:10.3892/or.2013.2316

Sorting, identification and enrichment of side population cells in THP-1 acute monocytic leukemia cells

Authors:
- Yingchao Wang
- Chuyun Yin
- Lei Feng
- Lina Ma
- Yongwei Wei
- Guangyao Sheng
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Affiliations: Department of Pediatrics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China
Published online on: February 28, 2013 https://doi.org/10.3892/or.2013.2316
Pages: 1923-1931

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Abstract

The objective of the present study was to examine and determine whether the human acute monocytic leukemia cell line THP-1 contains side population (SP) cells, and, if so, to increase the proportion of SP cells using arabinosylcytosine (Ara-C). Fluorescent microscopy and flow cytometry were employed to detect the percentage of SP cells in THP-1 cells. Then, SP and non-SP (NSP) cell subpopulations were collected and identified. THP-1 cells were incubated with different concentrations of Ara-C for 24 h and the proportion of SP cells was detected. Our 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 expression of CD34+ and CD34+CD38- and the proliferation ability of the SP cells were higher compared to NSP cells (P<0.05). The mRNA expression of multidrug resistance genes (ABCG2 and ABCB1), apoptosis regulation genes (Bcl-2) and the Bcl-2/Bax value of SP cells were higher than those of NSP cells. SP cells have been shown to be more tumorigenic than NSP cells. Following co-culture with Ara-C, the proportion of SP cells increased significantly and subsequently the Ara-C concentration increased. These ﬁndings suggest that the THP-1 cell line contains SP cells and that SP cells possess certain intrinsic stem cell properties and may contain a larger proportion of leukemia stem cells (LSCs). The concentrations of SP cells can be increased with Ara-C by co-culture, and this technique is a useful and important application for the study of LSCs.

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