TY - CHAP
T1 - Whole Transcriptome Sequencing Analysis of Cancer Stem/Progenitor Cells Obtained from Mouse Lung Adenocarcinomas
AU - Sinjab, Ansam
AU - Daouk, Reem
AU - Abou-Kheir, Wassim
AU - Kadara, Humam
N1 - Publisher Copyright:
© 2021, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2021
Y1 - 2021
N2 - Interrogation and characterization of lung cancer stem cells (CSCs) that are implicated in lung oncogenesis is crucial for our understanding of inter- and intra-tumor heterogeneity and the aggressive nature of the disease, including tumor resistance and relapse in response to conventional therapy. Here, we describe an in vitro surrogate model, namely the “sphere-forming assay,” for the derivation, enrichment, and propagation of lung stem/progenitor cells with CSC properties, including self-renewal, tumor initiation capacity and propagation, and differentiation into cells of the tumor bulk, from a murine Kras-mutant lung adenocarcinoma cell line. Self-renewing cancer stem/progenitor cells, in the form of 3D in vitro spheres, can be phenotypically interrogated using downstream techniques such as gene expression analysis (e.g., whole transcriptome sequencing and quantitative real time PCR), which is the focus of this chapter.
AB - Interrogation and characterization of lung cancer stem cells (CSCs) that are implicated in lung oncogenesis is crucial for our understanding of inter- and intra-tumor heterogeneity and the aggressive nature of the disease, including tumor resistance and relapse in response to conventional therapy. Here, we describe an in vitro surrogate model, namely the “sphere-forming assay,” for the derivation, enrichment, and propagation of lung stem/progenitor cells with CSC properties, including self-renewal, tumor initiation capacity and propagation, and differentiation into cells of the tumor bulk, from a murine Kras-mutant lung adenocarcinoma cell line. Self-renewing cancer stem/progenitor cells, in the form of 3D in vitro spheres, can be phenotypically interrogated using downstream techniques such as gene expression analysis (e.g., whole transcriptome sequencing and quantitative real time PCR), which is the focus of this chapter.
KW - 3D spheres
KW - Cancer stem/progenitor cells
KW - Quantitative real time PCR
KW - RNA-sequencing
KW - Self-renewal
UR - http://www.scopus.com/inward/record.url?scp=85102524949&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85102524949&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-1278-1_15
DO - 10.1007/978-1-0716-1278-1_15
M3 - Chapter
C2 - 33683695
AN - SCOPUS:85102524949
T3 - Methods in Molecular Biology
SP - 187
EP - 198
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -