Adaptation of Semiautomated Circulating Tumor Cell (CTC) Assays for Clinical and Preclinical Research Applications

Lori E. Lowes; Benjamin D. Hedley; Michael Keeney; Alison L. Allan

doi:10.3791/51248

Adaptación de semiautomatizado de células tumorales circulantes (CTC) Ensayos para aplicaciones c...

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Adaptation of Semiautomated Circulating Tumor Cell (CTC) Assays for Clinical and Preclinical Research Applications

Adaptación de semiautomatizado de células tumorales circulantes (CTC) Ensayos para aplicaciones clínicas y de investigación preclínica

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14:14 min

February 28, 2014

DOI: 10.3791/51248-v

Lori E. Lowes^1,2, Benjamin D. Hedley³, Michael Keeney^3,4, Alison L. Allan^1,2,4,5

¹London Regional Cancer Program,London Health Sciences Centre, ²Department of Anatomy & Cell Biology, Schulich School of Medicine and Dentistry,Western University, ³Special Hematology/Flow Cytometry,London Health Sciences Centre, ⁴Lawson Health Research Institute, ⁵Department of Oncology,Western University

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Overview

This study focuses on the detection, enumeration, and characterization of circulating tumor cells (CTCs) from human and mouse xenograft blood samples. It highlights the use of the CellSearch system for CTC analysis and describes protocols for user-defined marker characterization.

Key Study Components

Area of Science

Oncology
Cell Biology
Translational Research

Background

Circulating tumor cells (CTCs) serve as prognostic indicators in metastatic cancers.
The CellSearch system is the gold standard for CTC enumeration.
Common misclassification errors in CTC analysis are addressed.
Protocols for CTC characterization in preclinical models are developed.

Purpose of Study

To enhance the sensitivity of CTC detection and enumeration.
To characterize CTCs for better targeted therapy selection.
To validate the CellSearch system in both human and mouse models.

Methods Used

Collection of blood samples from human and mouse cancer models.
Positive selection for epca positive cells to enrich for CTCs.
Differential staining to identify and enumerate CTCs.
Molecular characterization of CTCs to inform therapy choices.

Main Results

CTCs were successfully detected and enumerated in both human and mouse samples.
The CellSearch system demonstrated effective CTC analysis.
User-defined protocols for CTC characterization were developed.
Results support the potential for improved targeted therapy selection.

Conclusions

The study confirms the efficacy of the CellSearch system for CTC analysis.
Adapted protocols may enhance CTC characterization for clinical applications.
Further development of these methods could aid in personalized cancer therapy.

Frequently Asked Questions

What are circulating tumor cells (CTCs)?

CTCs are cancer cells that have detached from the primary tumor and circulate in the bloodstream, serving as potential biomarkers for cancer progression.

How does the CellSearch system work?

The CellSearch system uses immunomagnetic separation and fluorescence microscopy to isolate and count CTCs from blood samples.

What is the significance of CTC enumeration?

CTC enumeration can provide prognostic information and help in monitoring treatment response in cancer patients.

Can CTCs be characterized for specific markers?

Yes, the study describes protocols for user-defined characterization of CTCs using specific markers.

What are the potential clinical applications of this research?

The findings may aid in the selection of targeted therapies for cancer patients based on CTC analysis.

Is this method applicable to both human and animal models?

Yes, the study demonstrates the effectiveness of the CellSearch system in both human and mouse xenograft blood samples.

Las células tumorales circulantes (CTC) son pronóstico en varios tipos de cáncer metastásico. Este manuscrito describe el sistema de patrón oro CellSearch (CSS) plataforma enumeración CTC y destaca los errores de clasificación errónea comunes. Además, dos protocolos adaptados se describen, por definido por el usuario caracterización marcador de CTCs y CTC enumeración en modelos de ratones preclínicos de metástasis que utilizan esta tecnología.

Este experimento tiene como objetivo la detección, enumeración y caracterización sensible de células tumorales circulantes a partir de muestras de sangre de xenoinjertos humanos y de ratón. En primer lugar, se recogen las muestras de sangre de modelos humanos y/o de ratón de cáncer y se utiliza una selección positiva para las células epca positivas que enriquecen para CTC en un contexto de leucocitos contaminantes. Luego, con la tinción diferencial de CTC y leucocitos, identifique y enumere las CTC.

Realizar la caracterización molecular deseada de estas células tumorales circulantes raras en el laboratorio para ayudar en la selección de terapias dirigidas para pacientes con cáncer en la clínica. Los resultados obtenidos utilizando el sistema de búsqueda celular demuestran que el análisis CTC se puede realizar de manera efectiva en muestras de sangre de xenoinjertos humanos y de ratón. El desarrollo de protocolos adicionales de caracterización de CTC definidos por el usuario en el sistema de búsqueda celular puede, en última instancia, ayudar en la selección de terapias dirigidas para los pacientes.

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