A modified method for isolation of bladder cancer stem cells from a MB49 murine cell line

BMC Urology

Research Article

Na Liu¹  Yong-jian Deng²  Wei Chen³  Yang Luo³  Yong-tong Zhu³  Wan-long Tan³  Fei Li³  Cheng-yong Lei³  Cheng-wu He³ 

[1] Department of Gynecology, Zhujiang Hospital, Southern Medical University, 510515, Guangzhou, Guangdong, P.R. China;Department of Pathology, Southern Medical University, 510515, Guangzhou, Guangdong, P.R. China;Department of Urology, Nanfang Hospital, Southern Medical University, 510515, Guangzhou, Guangdong, P.R. China;

关键词: Bladder cancer;    MB49 cell line;    Cancer stem cells;    Proliferation;    Chemotherapy;   

DOI  :  10.1186/1471-2490-13-57

 received in 2013-09-08, accepted in 2013-10-26,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundThe vaccine was efficiently effective against bladder cancer in earlier studies. However, a part of the mouse bladder tumour regrew due to regression after a period of time as the cancer stem cells could not be eliminated. In this study, we showed a modified method for the isolation of MB49 bladder cancer stem cells (MCSCs).MethodsThrough a comparison of different serum-free culture mediums (SFM), MCSCs were isolated by a combination of the limited dilution method and the optimal SFM method. The characterizations of MCSCs were verified by the fluorescence activated cell sorting, the quantitative polymerase chain reaction, the western blotting, the cell proliferation assay, the soft agar assay, the transwell assay, the resistance to chemotherapy assay and the tumor xenograft formation assay.ResultsThe optimal SFM contained a RPMI1640+ epidermal growth factor (20 ng/ml), a basic fibroblast growth factor (20 ng/ml), a leukemia inhibitory factor (20 ng/ml), a B-27 serum-free supplement (20 μl/ml), and a bovine serum albumin (4 μg/ml). MCSCs possessed the high expression of cancer stem cell markers (CD133, CD44, OCT4, NANOG, and ABCG2) and the ability of differentiation. In functional comparisons, MCSCs had higher proliferative abilities, lower susceptibility to chemotherapy, greater migration in vitro, and stronger tumorigenic abilities in vivo.ConclusionMCSCs displayed specific cancer stem cells properties. Our study showed MCSCs were isolated successfully with a modified method using a combination of limited dilution and SFM methods.

【 授权许可】

Unknown   
© Zhu et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.