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RNA-seq

RNA-seq

RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases.
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...

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Updated: Jun 1, 2025

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

Investigating subpopulation dynamics in clonal CHO-K1 cells with single-cell RNA sequencing.

Luke B Morina¹, Haoyu Chris Cao², Siqi Chen³

¹Johns Hopkins Biomedical Engineering, USA.

Journal of Biotechnology

|January 17, 2025

Summary

Investigating Chinese Hamster Ovary (CHO) cells reveals that seemingly identical clones harbor hidden cellular diversity. This heterogeneity, particularly the emergence of low-producing subpopulations, impacts biotherapeutic production stability.

Keywords:

CHO cells Clonal heterogeneity Single cell sequencing Transcriptomics

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

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Single-cell RNA Sequencing and Analysis of Human Pancreatic Islets

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Published on: July 18, 2019

Area of Science:

Biotechnology
Cell Biology
Biopharmaceutical Manufacturing

Background:

Chinese Hamster Ovary (CHO) cells are critical for industrial-scale biotherapeutic production.
Current understanding often overlooks cellular heterogeneity within ostensibly clonal CHO cell populations.
Assessing CHO cell behavior in bulk masks individual cell variations impacting production.

Purpose of the Study:

To investigate cellular heterogeneity in clonal CHO-K1 cell populations.
To understand the basis of variable stability phenotypes in long-term cultures.
To identify subpopulations contributing to reduced biotherapeutic production.

Main Methods:

Single-cell RNA sequencing was performed on two clonal CHO-K1 cell lines.
Cell populations were cultured over a 90-day period.
Gene expression profiles were analyzed to identify subpopulations.

Main Results:

One clonal CHO-K1 population exhibited instability due to a low-producing subpopulation.
This low-producing subpopulation emerged in aged cultures.
Low-producing cells did not show cellular stress markers present in high-producing cells.

Conclusions:

Cellular heterogeneity significantly impacts the stability of biotherapeutic production in CHO cells.
Emergence of distinct subpopulations contributes to decreased protein yield over time.
Further multiomic studies are needed to fully characterize CHO cell heterogeneity.