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Stem Cell Organoids in Primary Cultures of Human Non-Malignant and Malignant Colon

Authors :
Sahrish Tariq
Chris W. Sutton
Mariam Hassan
Mudassar Hussain
Muhammad Tahseen
Saira Saleem
Muhammed Aasim Yusuf
Shahid Khattak
Aamir Ali Syed
Asad Hayat Ahmad
Muhammad Adnan Masood
Source :
Journal of Molecular Biomarkers & Diagnosis.
Publication Year :
2017
Publisher :
OMICS Publishing Group, 2017.

Abstract

Aims: A sub-population of cells named cancer stem cells (CSCs) that initiate and promote tumour growth have been demonstrated to exist in several malignancies including colon carcinoma. The objective of our pilot study was to isolate CD133+CD26+CD44+ CSCs from patient colon tumours, culture spheres or organoids and observe their proliferation in primary cultures. Parallel cultures of non-cancer controls from colon normal lining and nonadenomatous polyps were set up. Methods: Magnetic activated cell sorting was used to isolate CD133+CD26+CD44+ cell populations followed by primary cell culturing under stem cell culture conditions. Number, cells/organoid and daughter generations of organoids were calculated using phase contrast microscope. Trypan blue exclusion method was used to test the viability of the cells. Results: Both colon tumour and colon non-adenomatous polyp formed floating organoids in suspension; however non-adenomatous polyp cultures did not show self-renewal properties for more than 1 passage. Normal colon singlecell suspension did not create organoids. Metastatic colon tumours rapidly produce cancer cell organoids in less than 24 hours in larger numbers compared to non-metastatic colon tumours (1-3 weeks). Metastatic colon tumour organoids have the ability for proliferation for upto five daughter generations in primary culture compared to three generations for those grown from non-metastatic tumours. Conclusions: This in vitro CSC organoid model will help study colon cancer biology, in particular providing a valuable source of primary cell-derived tissue for studying personalized molecular profiling using ‘omics strategies to direct therapeutic intervention.

Details

ISSN :
21559929
Database :
OpenAIRE
Journal :
Journal of Molecular Biomarkers & Diagnosis
Accession number :
edsair.doi...........014970905a6eb61f550957d282953000
Full Text :
https://doi.org/10.4172/2155-9929.1000346