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Serial Cell-free DNA Assessments in Preclinical Models
Ariana Rostami1,2, Caberry Yu1, Scott V Bratman1,2,3
1Princess Margaret Cancer Center, University Health Network, 101 College Street, Toronto, ON M5G 1L7, Canada.
STAR Protocols
|December 30, 2020
Summary
This study details a protocol for measuring circulating cell-free DNA (cfDNA) release in preclinical cancer models. The method allows for longitudinal evaluation of cfDNA dynamics during tumor development and treatment.
Area of Science:
- Biomedical research
- Molecular biology
- Cancer research
Background:
- Circulating cell-free DNA (cfDNA) is a valuable biomarker in oncology.
- Understanding cfDNA release mechanisms requires robust preclinical models.
- Longitudinal studies are crucial for tracking cfDNA dynamics.
Purpose of the Study:
- To present a detailed protocol for the longitudinal evaluation of cfDNA release.
- To establish a reproducible method for cfDNA analysis in preclinical cancer models.
- To facilitate mechanistic and kinetic investigations of cfDNA release.
Main Methods:
- Seeding cancer cell lines and establishing xenograft tumors.
- Treating cancer cells and xenograft tumors under various conditions.
- Serial collection of cell line media and xenograft blood for cfDNA isolation.
- Quantification of cfDNA using quantitative PCR (qPCR).
Main Results:
- The protocol enables serial cfDNA quantification.
- It allows for tracking cfDNA release kinetics over time.
- The method is applicable to both cell lines and in vivo xenografts.
Conclusions:
- The presented protocol provides a framework for studying cfDNA release.
- This method supports the investigation of cfDNA dynamics in cancer research.
- It aids in understanding the biological processes influencing cfDNA levels.
