Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
International Journal of Colorectal Disease
Published in: International Journal of Colorectal Disease 10/2011

01-10-2011 | Original Article

Isolation and characterization of spheroid cells from the HT29 colon cancer cell line

Authors: Xinlan Fan, Nengyong Ouyang, Hong Teng, Herui Yao

Published in: International Journal of Colorectal Disease | Issue 10/2011

Login to get access

Abstract

Background

Colorectal cancer stem cells (Cr-CSCs) are involved in the growth of colon cancer, but their specific role in tumor biology, including metastasis, is still unclear. Currently, methods for sorting Cr-CSCs are based on the expression of surface markers (e.g., CD133+, CD44+, and aldehyde dehydrogenase 1 (ALDH1+)); however, the specificity of these markers for Cr-CSCs is uncertain.

Purpose

This study aimed to develop more effective ways of isolating and purifying Cr-CSCs.

Methods

Suspension culture was used for isolation of Cr-CSCs. And spheroid cells were performed by side population technology, and the putative molecular marker analysis of colorectal cancer stem cell. Migration assay and chemoresistance experiment were conducted between the adherent cells and spheroid cells.

Results

HT29 colon cancer cells grew well in suspension culture. The percentage of CD44+ cancer cell of spheroid cells was 68 times higher than that of adherent cells (89.5% vs. 1.3%), but there was no obvious difference in the percentage of CD133+ cells (6.25% vs. 5.6%). Moreover, it is worth noting that the percent of CD133 +/CD44+ cells remarkably rose (from 0.6% to 5.4%). The expression of ALDH1 was markedly increased (7.5% vs. 20.5%) for the spheroid cells than the adherent cells. The side population within the spheroid population dramatically increased from 0.2% to 6.3%. The resistance of spheroid cells to 5-FU was higher than that of adherent cells, as was their ability to migrate in the presence of SDF-1α.

Conclusion

Suspension culture is an effective approach for enriching Cr-CSCs and can provide an inexhaustible supply of genetically stable colon cancer stem cells for targeted Cr-CSC studies. Spheroid cell models also enable the study of colon cancer chemoresistance and metastasis and may help to elucidate the role of cancer stem cells in colon cancer.
Literature
1.
Mittal S, Mifflin R, Powell DW (2009) Cancer stem cells: the other face of Janus. Am J Med Sc 338:107–112CrossRef
2.
Scopelliti A, Cammareri P, Catalano V et al (2009) Therapeutic implications of cancer initiating cells. Expert Opin Biol Ther 9:1005–1016PubMedCrossRef
3.
Ricci-Vitiani L, Lombardi DG, Pilozzi E et al (2007) Identification and expansion of human colon-cancer-initiating cells. Nature 445:111–115PubMedCrossRef
4.
O'Brien CA, Pollett A, Gallinger S et al (2007) A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 445:106–110PubMedCrossRef
5.
Shmelkov SV, Butler JM, Hooper AT et al (2008) CD133 expression is not restricted to stem cells, and both CD133+ and CD133- metastatic colon cancer cells initiate tumors. J Clin Invest 118:2111–2120PubMed
6.
Dalerba P, Dylla SJ, Park IK et al (2007) Phenotypic characterization of human colorectal cancer stem cells. Proc Natl Acad Sci 104:10158–10163PubMedCrossRef
7.
Huang EH, Hynes MJ, Zhang T et al (2009) Aldehyde dehydrogenase 1 is a marker for normal and malignant human colonic stem cells (SC) and tracks SC overpopulation during colon tumorigenesis. Cancer Res 69:3382–3389PubMedCrossRef
8.
Deng S, Yang X, Lassus H et al (2009) Distinct expression levels and patterns of stem cell marker, aldehyde dehydrogenase isoform 1 (ALDH1), in human epithelial cancers. PLoS One 5:e10277CrossRef
9.
Bar EE, Lin A, Mahairak V et al (2010) Hypoxia increases the expression of stem-cell markers and promotes clonogenicity in glioblastoma neurospheres. Am J Pathol 177:1491–1502PubMedCrossRef
10.
Rappa G, Lorico A (2010) Phenotypic characterization of mammosphere-forming cells from the human MA-11 breast carcinoma cell line. Exp Cell Res 316:1576–1586PubMedCrossRef
11.
Kanwar SS, Yu Y, Nautiyal J et al (2010) The Wnt/beta-catenin pathway regulates growth and maintenance of colonospheres. Mol Cancer 9:212–218PubMedCrossRef
12.
Goodell MA, Rosenzweig M, Kim H et al (1997) Dye efflux studies suggest that hematopoietic stem cells expressing low or undetectable levels of CD34 antigen exist in multiple species. Nat Med 3:1337–1345PubMedCrossRef
13.
Singh SK, Hawkins C, Clarke ID et al (2004) Identification of human brain tumour initiating cells. Nature 432:396–401PubMedCrossRef
14.
Dey D, Saxena M, Paranjape AN et al (2009) Phenotypic and functional characterization of human mammary stem/progenitor cells in long term culture. PLoS One 4:e5329PubMedCrossRef
15.
Todaro M, Alea MP, Di Stefano AB et al (2007) Colon cancer stem cells dictate tumor growth and resist cell death by production of interleukin-4. Cell Stem Cell 1:389–402PubMedCrossRef
16.
Yu F, Yao H, Zhu P et al (2007) let-7 regulates self renewal and tumorigenicity of breast cancer cells. Cell 131:1109–1123PubMedCrossRef
17.
Miller RJ, Banisadr G, Bhattacharyya BJ (2008) CXCR4 signaling in the regulation of stem cell migration and development. J Neuroimmunol 198:31–38PubMedCrossRef
18.
Hermann PC, Huber SL, Herrler T et al (2007) Distinct populations of cancer stem cells determines tumor growth and metastatic activity in human pancreatic cancer. Cell Stem Cell 1:313–323PubMedCrossRef
Metadata
Title
Isolation and characterization of spheroid cells from the HT29 colon cancer cell line
Authors
Xinlan Fan
Nengyong Ouyang
Hong Teng
Herui Yao
Publication date
01-10-2011
Publisher
Springer-Verlag
Published in
International Journal of Colorectal Disease / Issue 10/2011
Print ISSN: 0179-1958
Electronic ISSN: 1432-1262
DOI
https://doi.org/10.1007/s00384-011-1248-y

Other articles of this Issue 10/2011

International Journal of Colorectal Disease 10/2011 Go to the issue

Letter to the Editor

Pneumosalpynx caused by colosalpingeal fistula secondary to acute colonic diverticulitis

Review

Multiple mutations in the Kras gene in colorectal cancer: review of the literature with two case reports

Letter to the Editor

Sweeping KRAS generalizations: are we depriving patients of an effective treatment? A novel KRAS mutation and dramatic response to panitumumab in a patient with metastatic colorectal cancer

Original Article

Lack of association between XPD Lys751Gln and Asp312Asn polymorphisms and colorectal cancer risk: a meta-analysis of case–control studies

Letter to the Editor

Neoadjuvant chemoradiation therapy followed by curative transanal endoscopic microsurgery (TEM) for rectal cancer complicated by major suture dehiscence: avoiding ileostomy through hyperbaric oxygen therapy

Original Article

Cyclin D1 G870A polymorphism and colorectal cancer susceptibility: a meta-analysis of 20 populations