A marker-independent lineage-tracing system to quantify clonal dynamics and stem cell functionality in cancer tissue

Lenos, K. J., Lodestijn, S. C., Lyons, S. K., Bijlsma, M. F., Miedema, D. M., Vermeulen, L. (August 2019) A marker-independent lineage-tracing system to quantify clonal dynamics and stem cell functionality in cancer tissue. Nat Protoc, 14 (9). pp. 2648-2671. ISSN 1750-2799

URL: https://www.ncbi.nlm.nih.gov/pubmed/31420599
DOI: 10.1038/s41596-019-0194-y

Abstract

Lineage tracing is a powerful tool that can be used to uncover cell fates. Here, we describe a novel method for the quantitative analysis of clonal dynamics in grafted cancer tissues. The protocol involves the preparation and validation of cells for lineage tracing, establishment of grafts and label induction, analysis of clone-size distribution and fitting of the experimental data to a mathematical tumor growth model. In contrast to other lineage-tracing strategies, the method described here assesses stem cell functionality and infers tumor expansion dynamics independently of molecular markers such as putative cancer stem cell (CSC)-specific genes. The experimental system and analytical framework presented can be used to quantify clonal advantages that specific mutations provide, in both the absence and presence of (targeted) therapeutic agents. This protocol typically takes ~20 weeks to complete from cell line selection to inference of growth dynamics, depending on the grafted cancer growth rate.

Item Type: Paper
Subjects: diseases & disorders > cancer
diseases & disorders
Investigative techniques and equipment
diseases & disorders > neoplasms
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > cell line
Investigative techniques and equipment > cell lineage tracing
organs, tissues, organelles, cell types and functions > cell types and functions > cell types
organs, tissues, organelles, cell types and functions > cell types and functions > cell types
organs, tissues, organelles, cell types and functions > cell types and functions > cell types
organs, tissues, organelles, cell types and functions > cell types and functions
organs, tissues, organelles, cell types and functions
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > stem cells
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > stem cells
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > stem cells
CSHL Authors:
Communities: CSHL labs > Lyons lab
CSHL Cancer Center Program > Cellular Communication in Cancer Program
Depositing User: Matthew Dunn
Date: 16 August 2019
Date Deposited: 23 Aug 2019 18:23
Last Modified: 02 Feb 2024 16:44
Related URLs:
URI: https://repository.cshl.edu/id/eprint/38288

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