Isolation Set-up and Characterization of cancer stem cells from HT29 colon cancer cell line

Abstract Background: Cancer stem cells (CSCs) have critical roles in tumor initiation, progression, metastasis, resistance to chemoradiotherapy, and recurrence in various cancers such as colon cancer. Therefore, extensive studies are required to understand the CSCs mechanism of action and design nov...

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Autores principales:	Pooneh Mokarram, Mozhdeh Zamani, Marziyeh Babazadeh, Nasim Rahmani-Kukia, Masoomeh Rahimi, Seyed Vahid Hosseini
Formato:	article
Lenguaje:	EN
Publicado:	Shiraz University of Medical Sciences 2021
Materias:	cancer stem cell colon cancer stemness marker sphere formation Medicine R
Acceso en línea:	https://doaj.org/article/53bef5588f5d4871a72e2a93212dce46
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spelling	oai:doaj.org-article:53bef5588f5d4871a72e2a93212dce462021-11-14T06:50:55ZIsolation Set-up and Characterization of cancer stem cells from HT29 colon cancer cell line2783-243010.30476/acrr.2021.92388.1111https://doaj.org/article/53bef5588f5d4871a72e2a93212dce462021-09-01T00:00:00Zhttps://colorectalresearch.sums.ac.ir/article_47871_62329a44f56c2421d6a9cb6ee059c15a.pdfhttps://doaj.org/toc/2783-2430Abstract Background: Cancer stem cells (CSCs) have critical roles in tumor initiation, progression, metastasis, resistance to chemoradiotherapy, and recurrence in various cancers such as colon cancer. Therefore, extensive studies are required to understand the CSCs mechanism of action and design novel cancer therapies. Successful isolation and characterization of CSCs from tumor tissues or cancer cell lines provide the proper opportunity for these kinds of studies. This study aimed to isolate and characterize CSCs from HT29 colon cancer cell lines. Methods: Sphere formation assay was used to isolate CSCs from the HT29 cell line. The expression of four stemness genes, including LGR5, SOX2, c-Myc, and Oct4, was assessed by real-time PCR. The CSC markers, CD 44 and CD 24, were also evaluated and compared between the parental HT29 cell line and HT29-derived spheres by flow cytometry. Statistical analysis was performed using GraphPad Prism version 6.0 and the Mann-Whitney U test. Results:HT29-derived sphere cells were successfully formed in serum-free media. The proportion of CD 24 and CD 44-expressing cells and the expression of LGR5, SOX2, c-Myc, and Oct4 stemness genes were significantly higher in CSCs isolated from HT29 spheres compared with parental HT29 cells. Conclusion:Sphere formation assay is a proper method for the isolation of CSCs from the HT29 cell line. The stemness markers LGR5, SOX2, c-Myc, Oct4,CD 44,andCD 24are suitable for the characterization of these cells.Pooneh MokarramMozhdeh ZamaniMarziyeh BabazadehNasim Rahmani-KukiaMasoomeh RahimiSeyed Vahid HosseiniShiraz University of Medical Sciencesarticlecancer stem cellcolon cancerstemness markersphere formationMedicineRENIranian Journal of Colorectal Research, Vol 9, Iss 3, Pp 103-108 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	cancer stem cell colon cancer stemness marker sphere formation Medicine R
spellingShingle	cancer stem cell colon cancer stemness marker sphere formation Medicine R Pooneh Mokarram Mozhdeh Zamani Marziyeh Babazadeh Nasim Rahmani-Kukia Masoomeh Rahimi Seyed Vahid Hosseini Isolation Set-up and Characterization of cancer stem cells from HT29 colon cancer cell line
description	Abstract Background: Cancer stem cells (CSCs) have critical roles in tumor initiation, progression, metastasis, resistance to chemoradiotherapy, and recurrence in various cancers such as colon cancer. Therefore, extensive studies are required to understand the CSCs mechanism of action and design novel cancer therapies. Successful isolation and characterization of CSCs from tumor tissues or cancer cell lines provide the proper opportunity for these kinds of studies. This study aimed to isolate and characterize CSCs from HT29 colon cancer cell lines. Methods: Sphere formation assay was used to isolate CSCs from the HT29 cell line. The expression of four stemness genes, including LGR5, SOX2, c-Myc, and Oct4, was assessed by real-time PCR. The CSC markers, CD 44 and CD 24, were also evaluated and compared between the parental HT29 cell line and HT29-derived spheres by flow cytometry. Statistical analysis was performed using GraphPad Prism version 6.0 and the Mann-Whitney U test. Results:HT29-derived sphere cells were successfully formed in serum-free media. The proportion of CD 24 and CD 44-expressing cells and the expression of LGR5, SOX2, c-Myc, and Oct4 stemness genes were significantly higher in CSCs isolated from HT29 spheres compared with parental HT29 cells. Conclusion:Sphere formation assay is a proper method for the isolation of CSCs from the HT29 cell line. The stemness markers LGR5, SOX2, c-Myc, Oct4,CD 44,andCD 24are suitable for the characterization of these cells.
format	article
author	Pooneh Mokarram Mozhdeh Zamani Marziyeh Babazadeh Nasim Rahmani-Kukia Masoomeh Rahimi Seyed Vahid Hosseini
author_facet	Pooneh Mokarram Mozhdeh Zamani Marziyeh Babazadeh Nasim Rahmani-Kukia Masoomeh Rahimi Seyed Vahid Hosseini
author_sort	Pooneh Mokarram
title	Isolation Set-up and Characterization of cancer stem cells from HT29 colon cancer cell line
title_short	Isolation Set-up and Characterization of cancer stem cells from HT29 colon cancer cell line
title_full	Isolation Set-up and Characterization of cancer stem cells from HT29 colon cancer cell line
title_fullStr	Isolation Set-up and Characterization of cancer stem cells from HT29 colon cancer cell line
title_full_unstemmed	Isolation Set-up and Characterization of cancer stem cells from HT29 colon cancer cell line
title_sort	isolation set-up and characterization of cancer stem cells from ht29 colon cancer cell line
publisher	Shiraz University of Medical Sciences
publishDate	2021
url	https://doaj.org/article/53bef5588f5d4871a72e2a93212dce46
work_keys_str_mv	AT poonehmokarram isolationsetupandcharacterizationofcancerstemcellsfromht29coloncancercellline AT mozhdehzamani isolationsetupandcharacterizationofcancerstemcellsfromht29coloncancercellline AT marziyehbabazadeh isolationsetupandcharacterizationofcancerstemcellsfromht29coloncancercellline AT nasimrahmanikukia isolationsetupandcharacterizationofcancerstemcellsfromht29coloncancercellline AT masoomehrahimi isolationsetupandcharacterizationofcancerstemcellsfromht29coloncancercellline AT seyedvahidhosseini isolationsetupandcharacterizationofcancerstemcellsfromht29coloncancercellline
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Isolation Set-up and Characterization of cancer stem cells from HT29 colon cancer cell line

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