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A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

Published on: October 25, 2018

Cellular heterogeneity in the 16HBE14o

Jenny L Kerschner¹, Alekh Paranjapye¹, Ann Harris¹

¹Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, Ohio, USA.

Physiological Reports

|June 3, 2023

Summary

Heterogeneity in airway epithelial cells (16HBE14o-) affects cystic fibrosis transmembrane conductance regulator (CFTR) expression. Clonal variations impact immune responses, urging caution when interpreting data from manipulated cell lines.

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Area of Science:

Cell Biology
Genomics
Immunology

Background:

The 16HBE14o- cell line is crucial for airway disease research.
SV40-mediated immortalization can lead to genomic instability in long-term cultured cells.
Variations in gene expression, such as for CFTR, can arise within cell line clones.

Purpose of the Study:

To investigate heterogeneity in 16HBE14o- cells regarding CFTR expression.
To characterize CFTR locus accessibility and chromatin structure in relation to CFTR levels.
To identify transcriptomic differences, including immune responses, between CFTR-expressing clones.

Main Methods:

Isolation and characterization of CFTR-high and CFTR-low 16HBE14o- cell clones.
ATAC-seq and 4C-seq to analyze chromatin accessibility and structure at the CFTR locus.
RNA-sequencing to compare transcriptomes of CFTR-high and CFTR-low cells.

Main Results:

Stable CFTR-high and CFTR-low clones were successfully isolated from 16HBE14o- cells.
Chromatin profiles at the CFTR locus correlated with CFTR expression levels.
CFTR-high cells exhibited an enhanced inflammatory and innate immune response phenotype.

Conclusions:

Significant heterogeneity exists within the 16HBE14o- cell line concerning CFTR expression.
Chromatin structure plays a role in regulating CFTR expression in these cells.
Caution is advised when interpreting functional data from manipulated 16HBE14o- clonal lines due to potential immune response alterations.