Serial Cell-free DNA Assessments in Preclinical Models

• Published in: STAR protocols 2020-12, Vol.1 (3), p.100145-100145, Article 100145
• Main Authors: Rostami, Ariana, Yu, Caberry, Bratman, Scott V.
• Format: Article
• Language: English
• Subjects: Animals; Cell Line, Tumor; Cell-Free Nucleic Acids - analysis; Cell-Free Nucleic Acids - genetics; Culture Media - analysis; Humans; Mice; Protocol; Real-Time Polymerase Chain Reaction - methods; Xenograft Model Antitumor Assays - methods
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2020.100145

Studying circulating cell-free DNA (cfDNA) release within preclinical model systems provides opportunities to investigate the mechanisms and kinetics underlying this process under various conditions. We present a detailed protocol for longitudinal evaluation of cfDNA release through (1) seeding of cancer cell lines and establishment of xenograft tumors, (2) treatment of cancer cells and xenograft tumors, (3) serial collection of cell line media and xenograft blood, and (4) processing and isolation of cfDNA for (5) quantification of cfDNA by quantitative PCR. For complete details on the use and execution of this protocol please refer to Rostami et al. (2020). [Display omitted] •Cell line media and xenograft blood can be serially collected after irradiation•Small plasma volumes can be simultaneously purified for cell-free DNA (cfDNA)•Cell line media can be diluted for quantification of cfDNA using quantitative PCR•Human LINE-1 can be used for ultrasensitive detection of cfDNA Studying circulating cell-free DNA (cfDNA) release within preclinical model systems provides opportunities to investigate the mechanisms and kinetics underlying this process under various conditions. We present a detailed protocol for longitudinal evaluation of cfDNA release through (1) seeding of cancer cell lines and establishment of xenograft tumors, (2) treatment of cancer cells and xenograft tumors, (3) serial collection of cell line media and xenograft blood, and (4) processing and isolation of cfDNA for (5) quantification of cfDNA by quantitative PCR.