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Published on: June 1, 2018
Serum-Based Culture Conditions Provoke Gene Expression Variability in Mouse Embryonic Stem Cells as Revealed by
Guoji Guo1, Luca Pinello2, Xiaoping Han1
1Division of Pediatric Hematology/Oncology, Dana Farber Cancer Institute and Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA; Center for Stem Cell and Regenerative Medicine, Zhejiang University School of Medicine, Hangzhou 310058, China.
Gene expression varies significantly in mouse embryonic stem cells (ESCs). Serum-free culture reduces this cellular heterogeneity and transcriptome variation, revealing the impact of the environment on ESCs.
Area of Science:
- Stem cell biology
- Transcriptomics
- Epigenetics
Background:
- Gene expression variation is a key feature of mouse embryonic stem cells (ESCs).
- Mechanisms driving global gene expression variability in ESCs remain incompletely understood.
- Understanding ESC heterogeneity is crucial for developmental biology and regenerative medicine.
Purpose of the Study:
- To investigate the mechanisms underlying global gene expression variation in mouse ESCs.
- To identify factors contributing to cellular heterogeneity within ESC populations.
- To explore the impact of culture environment on ESC transcriptome variation.
Main Methods:
- Single-cell messenger RNA sequencing (mRNA-seq) was employed on mouse ESCs.
- Analysis of gene expression patterns, chromatin states, and bivalent gene regulation.
- Comparison of ESCs cultured in serum-containing versus serum-free conditions.
Main Results:
- Significant cellular heterogeneity and transcriptome variation were observed in ESCs cultured in serum.
- Highly variable gene clusters with distinct chromatin states were identified.
- Bivalent genes were found to be particularly prone to expression variation.
- An ESC-priming pathway initiating exit from the naive state was identified.
- The extracellular culture environment was identified as a major contributor to intracellular network variability.
- Serum-free culture conditions significantly reduced cellular heterogeneity and transcriptome variation in ESCs.
Conclusions:
- The culture environment, particularly serum, plays a substantial role in driving gene expression variation and heterogeneity in ESCs.
- Serum-free culture represents a viable strategy to minimize transcriptome variation and maintain ESC homogeneity.
- These findings provide insights into ESC regulation and offer practical implications for stem cell culture and applications.
