Isolation of Breast Cancer Stem Cell from MDA-MB231 Cell Line Using Vincristine

Cancer has been considered as a stem cell disease. Suspension culture combined with anti-cancer drugs has recently been proposed for isolation of cancer stem cells (CSCs). In the current study, Vincristine as an anti-cancer drug combined with suspension culture was used for isolation and purificatio...

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Auteurs principaux:	Ghanbari,Tara, Azadbakht,Mehri, Vesi-Raygani,Asad, Khazaei,Mozafar
Langue:	English
Publié:	Sociedad Chilena de Anatomía 2016
Sujets:	Cancer stem cell Vincristine Suspension culture MDA-MB231
Accès en ligne:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022016000400003
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spelling	oai:scielo:S0717-950220160004000032017-02-01Isolation of Breast Cancer Stem Cell from MDA-MB231 Cell Line Using VincristineGhanbari,TaraAzadbakht,MehriVesi-Raygani,AsadKhazaei,Mozafar Cancer stem cell Vincristine Suspension culture MDA-MB231 Cancer has been considered as a stem cell disease. Suspension culture combined with anti-cancer drugs has recently been proposed for isolation of cancer stem cells (CSCs). In the current study, Vincristine as an anti-cancer drug combined with suspension culture was used for isolation and purification of CSCs from human breast cancer cell line (MDA-MB231). The cells were treated with different concentrations of vincristine (0, 2, 4, 6 and 8 ng/ml). Stem cells were identified with the expression of OCT4, nanog, SOX2 and nucleostemin genes by RT-PCR. Mammosphere forming unit was measured upon suspension culture containing EGF, bFGF, LIF, B27, insulin and BSA. The isolated mammospheres were investigated for CD44 expression. Results showed that 4 ng/ml of vincristine for 72 hours could be utilized as the best and most reliable dose which eliminates around 80 % of non-cancer stem cells with no destructive effect on CSCs' viability (P> 0.05). RT-PCR demonstrated that drug treated cells expressed OCT4, nanog, SOX2 and nucleostemin. Mammosphere formation unit of cells pretreated with vincristine was significantly higher than unpretreated ones (P>0.05). Immunofluorescence staining for CD44 depicted high expression of CSC marker among the isolated mammospheres. Vincristine combined with suspension culture can be considered as an appropriate method to isolate CSC.info:eu-repo/semantics/openAccessSociedad Chilena de AnatomíaInternational Journal of Morphology v.34 n.4 20162016-12-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022016000400003en10.4067/S0717-95022016000400003
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Cancer stem cell Vincristine Suspension culture MDA-MB231
spellingShingle	Cancer stem cell Vincristine Suspension culture MDA-MB231 Ghanbari,Tara Azadbakht,Mehri Vesi-Raygani,Asad Khazaei,Mozafar Isolation of Breast Cancer Stem Cell from MDA-MB231 Cell Line Using Vincristine
description	Cancer has been considered as a stem cell disease. Suspension culture combined with anti-cancer drugs has recently been proposed for isolation of cancer stem cells (CSCs). In the current study, Vincristine as an anti-cancer drug combined with suspension culture was used for isolation and purification of CSCs from human breast cancer cell line (MDA-MB231). The cells were treated with different concentrations of vincristine (0, 2, 4, 6 and 8 ng/ml). Stem cells were identified with the expression of OCT4, nanog, SOX2 and nucleostemin genes by RT-PCR. Mammosphere forming unit was measured upon suspension culture containing EGF, bFGF, LIF, B27, insulin and BSA. The isolated mammospheres were investigated for CD44 expression. Results showed that 4 ng/ml of vincristine for 72 hours could be utilized as the best and most reliable dose which eliminates around 80 % of non-cancer stem cells with no destructive effect on CSCs' viability (P> 0.05). RT-PCR demonstrated that drug treated cells expressed OCT4, nanog, SOX2 and nucleostemin. Mammosphere formation unit of cells pretreated with vincristine was significantly higher than unpretreated ones (P>0.05). Immunofluorescence staining for CD44 depicted high expression of CSC marker among the isolated mammospheres. Vincristine combined with suspension culture can be considered as an appropriate method to isolate CSC.
author	Ghanbari,Tara Azadbakht,Mehri Vesi-Raygani,Asad Khazaei,Mozafar
author_facet	Ghanbari,Tara Azadbakht,Mehri Vesi-Raygani,Asad Khazaei,Mozafar
author_sort	Ghanbari,Tara
title	Isolation of Breast Cancer Stem Cell from MDA-MB231 Cell Line Using Vincristine
title_short	Isolation of Breast Cancer Stem Cell from MDA-MB231 Cell Line Using Vincristine
title_full	Isolation of Breast Cancer Stem Cell from MDA-MB231 Cell Line Using Vincristine
title_fullStr	Isolation of Breast Cancer Stem Cell from MDA-MB231 Cell Line Using Vincristine
title_full_unstemmed	Isolation of Breast Cancer Stem Cell from MDA-MB231 Cell Line Using Vincristine
title_sort	isolation of breast cancer stem cell from mda-mb231 cell line using vincristine
publisher	Sociedad Chilena de Anatomía
publishDate	2016
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022016000400003
work_keys_str_mv	AT ghanbaritara isolationofbreastcancerstemcellfrommdamb231celllineusingvincristine AT azadbakhtmehri isolationofbreastcancerstemcellfrommdamb231celllineusingvincristine AT vesirayganiasad isolationofbreastcancerstemcellfrommdamb231celllineusingvincristine AT khazaeimozafar isolationofbreastcancerstemcellfrommdamb231celllineusingvincristine
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Isolation of Breast Cancer Stem Cell from MDA-MB231 Cell Line Using Vincristine

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