Isolation and Propagation of Colon Cancer Stem Cells In vit

Document Type : Original Article (s)

Authors

1 Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan

2 Assistant Professor, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 Professor, Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

4 Associate Professor, Department of Internal Medicine, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

5 Associate Professor, Department of Pathology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

6 Associate Professor, Department of Biostatistics and Epidemiology, School of Public Health, Isfahan University of Medical Sciences, Isfahan, Iran

7 Assistant Professor, Department of Surgery, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

8 Associate Professor, Department of Surgery, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Recent investigations in colon cancer biology suggest that cancer growth is driven by colon cancer stem cells (CSCs). CSCs are responsible for tumor initiation, maintenance, spreading and relapse. The aim of this study was to isolate and propagate tumorigenic colon CSCs in vitro.Methods: Tumor samples from colon cancers were subjected to mechanical and enzymatic disassociation. After passing of tumor cell suspensions through the cell strainers, tumor cells were counted and their viability was determined. Then tumor cells were plated in the special stem cells medium (contains serum-free DMEM/F12 supplemented by growth factors). Tumor cells growth was evaluated by inverted microscope.Findings: After about 3 weeks of plating in specific medium, colon CSCs were appeared as cell spheres whereas differentiated tumor cells and nonmalaginent cells were unable to grow. Colon cancer spheres were able to propagated and passaged in the specific stem cells medium.Conclusion: Isolation and propagation of tumorigenic colon CSCs in vitro would help to devise novel diagnostic and therapeutic methods. Isolated colon CSCs can be applied for the studying of cell signaling and assessment of the effect of anti-cancer drugs.

Keywords

References

  1. Foroutan M, Rahimi N, Tabatabaeifar M, Darvishi M, Hashemi M, Hossein-Panah F, et al. Clinical features of colorectal cancer in Iran: a 15-year review. J Dig Dis 2008; 9(4): 225-7.
  2. Singh SK, Hawkins C, Clarke ID, Squire JA, Bayani J, Hide T, et al. Identification of human brain tumour initiating cells. Nature 2004; 432(7015): 396-401.
  3. Ricci-Vitiani L, Lombardi DG, Pilozzi E, Biffoni M, Todaro M, Peschle C, et al. Identification and expansion of human colon-cancer-initiating cells. Nature 2007; 445(7123): 111-5.
  4. O'Brien CA, Pollett A, Gallinger S, Dick JE. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 2007; 445(7123): 106-10.
  5. Fang D, Nguyen TK, Leishear K, Finko R, Kulp AN, Hotz S, et al. A tumorigenic subpopulation with stem cell properties in melanomas. Cancer Res 2005; 65(20): 9328-37.
  6. Fillmore CM, Kuperwasser C. Human breast cancer cell lines contain stem-like cells that self-renew, give rise to phenotypically diverse progeny and survive chemotherapy. Breast Cancer Res 2008; 10(2): R25.
  7. Ricci-Vitiani L, Fabrizi E, Palio E, De MR. Colon cancer stem cells. J Mol Med (Berl) 2009; 87(11): 1097-104.
  8. Todaro M, Perez AM, Scopelliti A, Medema JP, Stassi G. IL-4-mediated drug resistance in colon cancer stem cells. Cell Cycle 2008; 7(3): 309-13.
  9. Croker AK, Allan AL. Inhibition of aldehyde dehydrogenase (ALDH) activity reduces chemotherapy and radiation resistance of stem-like ALDH(hi)CD44 (+) human breast cancer cells. Breast Cancer Res Treat 2011.
  10. Pajonk F, Vlashi E, McBride WH. Radiation resistance of cancer stem cells: the 4 R's of radiobiology revisited. Stem Cells 2010; 28(4): 639-48.
  11. Debeb BG, Xu W, Woodward WA. Radiation resistance of breast cancer stem cells: understanding the clinical framework. J Mammary Gland Biol Neoplasia 2009; 14(1): 11-7.
  12. Dylla SJ, Beviglia L, Park IK, Chartier C, Raval J, Ngan L, et al. Colorectal cancer stem cells are enriched in xenogeneic tumors following chemotherapy. PLoS One 2008; 3(6): e2428.
  13. Pang R, Law WL, Chu AC, Poon JT, Lam CS, Chow AK, et al. A subpopulation of CD26+ cancer stem cells with metastatic capacity in human colorectal cancer. Cell Stem Cell 2010; 6(6): 603-15.
  14. Bao S, Wu Q, McLendon RE, Hao Y, Shi Q, Hjelmeland AB, et al. Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature 2006; 444(7120): 756-60.
  15. Shmelkov SV, St CR, Lyden D, Rafii S. AC133/CD133/Prominin-1. Int J Biochem Cell Biol 2005; 37(4): 715-9.
  16. Shmelkov SV, Butler JM, Hooper AT, Hormigo A, Kushner J, Milde T, et al. CD133 expression is not restricted to stem cells, and both CD133+ and C. J Clin Invest 2008; 118(6): 2111-20.
  17. Du L, Wang H, He L, Zhang J, Ni B, Wang X, et al. CD44 is of functional importance for colorectal cancer stem cells. Clin Cancer Res 2008; 14(21): 6751-60.
  18. Dalerba P, Dylla SJ, Park IK, Liu R, Wang X, Cho RW, et al. Phenotypic characterization of human colorectal cancer stem cells. Proc Natl Acad Sci U S A 2007; 104(24): 10158-63.
Journal of Isfahan Medical School
Volume 29, Issue 154 - Serial Number 154
July and August 2011
Pages 1356-1364

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History

  • Receive Date: 05 October 2011
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

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