A modified method by differential adhesion for enrichment of bladder cancer stem cells

ABSTRACT Purpose: In a previous study the vaccine was effective against bladder cancer in a mouse model. However, a small portion of tumors regrew because the vaccine could not eliminate bladder cancer stem cells (CSCs). In this study, we showed a modified method for the isolation of bladder CSCs using a combination of differential adhesion method and serum-free culture medium (SFM) method. Materials and Methods: Trypsin-resistant cells and trypsin-sensitive cells were isolated from MB49, EJ and 5637 cells by a combination of differential adhesion method and SFM method. The CSCs characterizations of trypsin-resistant cells were verified by the flow cytometry, the western blotting, the quantitative polymerase chain reaction, the resistance to chemotherapy assay, the transwell assay, and the tumor xenograft formation assay. Results: Trypsin-resistant cells were isolated and identified in CSCs characters, with high expression of CSCs markers, higher resistance to chemotherapy, greater migration in vitro, and stronger tumorigenicity in vivo. Conclusion: Trypsin-resistant cells displayed specific CSCs properties. Our study showed trypsin-resistant cells were isolated successfully with a modified method using a combination of differential adhesion method and SFM method.

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Main Authors: Zhu,Yong-tong, Pang,Shi-yu, Luo,Yang, Chen,Wei, Bao,Ji-ming, Tan,Wan-long
Format: Digital revista
Language:English
Published: Sociedade Brasileira de Urologia 2016
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1677-55382016000400817
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spelling oai:scielo:S1677-553820160004008172016-09-06A modified method by differential adhesion for enrichment of bladder cancer stem cellsZhu,Yong-tongPang,Shi-yuLuo,YangChen,WeiBao,Ji-mingTan,Wan-long Urinary Bladder Neoplasms Neoplastic Stem Cells Trypsin ABSTRACT Purpose: In a previous study the vaccine was effective against bladder cancer in a mouse model. However, a small portion of tumors regrew because the vaccine could not eliminate bladder cancer stem cells (CSCs). In this study, we showed a modified method for the isolation of bladder CSCs using a combination of differential adhesion method and serum-free culture medium (SFM) method. Materials and Methods: Trypsin-resistant cells and trypsin-sensitive cells were isolated from MB49, EJ and 5637 cells by a combination of differential adhesion method and SFM method. The CSCs characterizations of trypsin-resistant cells were verified by the flow cytometry, the western blotting, the quantitative polymerase chain reaction, the resistance to chemotherapy assay, the transwell assay, and the tumor xenograft formation assay. Results: Trypsin-resistant cells were isolated and identified in CSCs characters, with high expression of CSCs markers, higher resistance to chemotherapy, greater migration in vitro, and stronger tumorigenicity in vivo. Conclusion: Trypsin-resistant cells displayed specific CSCs properties. Our study showed trypsin-resistant cells were isolated successfully with a modified method using a combination of differential adhesion method and SFM method.info:eu-repo/semantics/openAccessSociedade Brasileira de UrologiaInternational braz j urol v.42 n.4 20162016-08-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1677-55382016000400817en10.1590/S1677-5538.IBJU.2015.0409
institution SCIELO
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country Brasil
countrycode BR
component Revista
access En linea
databasecode rev-scielo-br
tag revista
region America del Sur
libraryname SciELO
language English
format Digital
author Zhu,Yong-tong
Pang,Shi-yu
Luo,Yang
Chen,Wei
Bao,Ji-ming
Tan,Wan-long
spellingShingle Zhu,Yong-tong
Pang,Shi-yu
Luo,Yang
Chen,Wei
Bao,Ji-ming
Tan,Wan-long
A modified method by differential adhesion for enrichment of bladder cancer stem cells
author_facet Zhu,Yong-tong
Pang,Shi-yu
Luo,Yang
Chen,Wei
Bao,Ji-ming
Tan,Wan-long
author_sort Zhu,Yong-tong
title A modified method by differential adhesion for enrichment of bladder cancer stem cells
title_short A modified method by differential adhesion for enrichment of bladder cancer stem cells
title_full A modified method by differential adhesion for enrichment of bladder cancer stem cells
title_fullStr A modified method by differential adhesion for enrichment of bladder cancer stem cells
title_full_unstemmed A modified method by differential adhesion for enrichment of bladder cancer stem cells
title_sort modified method by differential adhesion for enrichment of bladder cancer stem cells
description ABSTRACT Purpose: In a previous study the vaccine was effective against bladder cancer in a mouse model. However, a small portion of tumors regrew because the vaccine could not eliminate bladder cancer stem cells (CSCs). In this study, we showed a modified method for the isolation of bladder CSCs using a combination of differential adhesion method and serum-free culture medium (SFM) method. Materials and Methods: Trypsin-resistant cells and trypsin-sensitive cells were isolated from MB49, EJ and 5637 cells by a combination of differential adhesion method and SFM method. The CSCs characterizations of trypsin-resistant cells were verified by the flow cytometry, the western blotting, the quantitative polymerase chain reaction, the resistance to chemotherapy assay, the transwell assay, and the tumor xenograft formation assay. Results: Trypsin-resistant cells were isolated and identified in CSCs characters, with high expression of CSCs markers, higher resistance to chemotherapy, greater migration in vitro, and stronger tumorigenicity in vivo. Conclusion: Trypsin-resistant cells displayed specific CSCs properties. Our study showed trypsin-resistant cells were isolated successfully with a modified method using a combination of differential adhesion method and SFM method.
publisher Sociedade Brasileira de Urologia
publishDate 2016
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1677-55382016000400817
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