Size-based method for enrichment of circulating tumor cells from blood of colorectal cancer patients
Abstract
Circulating tumor cells (CTCs) are precursors of the metastatic cascade, which is responsible for 90% of all cancer-related deaths. CTCs arise from solid tumors and travel through the bloodstream and lymphatic system. Developments in the isolation and analysis of CTCs promise potential biomarker assays for detection and monitoring of cancer through a minimally invasive procedure. Despite this, the precise role of CTCs in metastasis remains poorly characterized, mainly due to the low density of CTCs (1-10 CTCs per 10 mL of blood) present in patient blood and the lack of robust methods for their isolation in a standard laboratory setting. CellSearch is currently the only FDA-approved CTC enrichment protocol, but limitations of this EpCAM-based method include cost, availability, and the use of a single surface marker for capture. To address these limitations, we have optimized an existing method, MetaCell, which exploits the differences in size of CTCs compared to other blood cells for CTC enrichment from blood. MetaCell contains a membrane with 8 μm pores, and blood is filtered through this kit by capillary action and CTCs, which are typically larger than the pores and remain on top of the membrane, while most leukocytes pass through the pores. The membrane along with these CTCs can be detached and transferred to 6-well plates for culturing or directly used for characterization. Here, we provide a detailed protocol for enrichment of CTCs using the filtration device MetaCell and a procedure for characterization of CTCs by immunohistochemical staining.
RAS ID
55467
Document Type
Book Chapter
Date of Publication
1-1-2023
Volume
2588
Funding Information
Rutherford Discovery fellowship Ministry of Business, Innovation and Employment (MBIE), New Zealand
PubMed ID
36418692
School
Centre for Precision Health / School of Medical and Health Sciences
Copyright
subscription content
Publisher
Springer
Recommended Citation
Vasantharajan, S. S., Barnett, E., Gray, E. S., Rodger, E. J., Eccles, M. R., Pattison, S., Munro, F., & Chatterjee, A. (2023). Size-based method for enrichment of circulating tumor cells from blood of colorectal cancer patients. DOI: https://doi.org/10.1007/978-1-0716-2780-8_15
Comments
Vasantharajan, S. S., Barnett, E., Gray, E. S., Rodger, E. J., Eccles, M. R., Pattison, S., . . . Chatterjee, A. (2023). Size-based method for enrichment of circulating tumor cells from blood of colorectal cancer patients. In G. J. Seymour, M. P. Cullinan, N. C. K. Henag & P. R. Cooper (Eds.), Oral Biology (pp. 231-248). Springer. https://doi.org/10.1007/978-1-0716-2780-8_15