Isolation and enrichment of PC-3 prostate cancer stem-like cells using MACS and serum-free medium

Sheng,Xia; Li,Zeng; Wang,De-Lin; Li,Wen-Bin; Luo,Zhao; Chen,Ke-Hong; Cao,Jian-Jia; Yu,Chao; Liu,Wu-Jiang

doi:10.3892/ol.2012.1090

Isolation and enrichment of PC-3 prostate cancer stem-like cells using MACS and serum-free medium

Authors:
- Xia Sheng
- Zeng Li
- De-Lin Wang
- Wen-Bin Li
- Zhao Luo
- Ke-Hong Chen
- Jian-Jia Cao
- Chao Yu
- Wu-Jiang Liu
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Affiliations: Department of Urology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China, Life Science Research Institute of Chongqing Medical University, Chongqing 400016, P.R. China, Institute of Urology, Peking University First Hospital Street 8 of Xishiku, Xicheng, Beijing 100034, P.R. China
Published online on: December 21, 2012 https://doi.org/10.3892/ol.2012.1090
Pages: 787-792

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Abstract

Prostate cancer stem-like cells (PCSLCs) are considered to be the ‘seed’ of prostate cancer. The aim of this study was to confirm that the PC‑3 cells, which we isolated and enriched from PC-3 cells through magnetic bead cell sorting (MACS) and serum-free medium (SFM) culture, were PCSLCs. Combinations of MACS, flow cytometry (FCM), SFM and immunocytochemistry (ICC) were used to ensure the positive expression of CD133 and CD44 on PC‑3 and sphere‑forming cell membranes. Self-renewal, multi-potential differentiation, unlimited proliferation and permanency assays were also applied to indentify whether the PC‑3 cells exhibited the characteristics of cancer stem cells (CSCs). As a result, there was a low proportion of PCSLCs in the PC-3 cells. In the FCM assay, the proportion of cells expressing CD133 or CD44 in the PC-3 cells was 0.51 and 0.31%, respectively. In addition, we found that the proportion of PC‑3 cells sorted by MACS that expressed CD133 was significantly increased compared with that of the sphere‑forming cells cultured in SFM (99.09 vs. 84.80%, P<0.05), while no difference was observed in the proportion of cells expressing CD44 between them (99.88 vs. 99.82%, P>0.05). The expression of PAP and AR as detected by western blot analysis of induced PCSLCs was significantly increased compared with that of uninduced PCSLCs (P<0.05); the proliferation capacity of PCSLCs was significantly higher than that of both the PC-3 cells (P<0.05) and induced PCSLCs (P<0.05). Furthermore, the PCSLCs that were isolated from SFM and MACS both demonstrated certain characteristics of stem cells and should be considered as stem cell‑like. These data may hold potential for further exploring the role of PCSLCs.

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