Transcriptome analysis of gene expression changes upon enzymatic dissociation in skeletal myoblasts

Area of Science:

• Molecular Biology
• Genomics
• Cell Biology

Background:

• Single-cell RNA sequencing (scRNA-seq) is crucial for understanding cellular heterogeneity.
• Cell isolation via enzymatic dissociation is a prerequisite for scRNA-seq.
• The impact of enzymatic dissociation on gene expression profiles remains incompletely understood.

Purpose of the Study:

• To comprehensively profile enzyme-dependent gene expression changes during cell dissociation.
• To investigate the influence of enzyme treatment time, concentration, and temperature.
• To provide reference data for optimizing RNA-seq protocols.

Main Methods:

• Mouse skeletal muscle progenitor cells were subjected to enzymatic dissociation using varying conditions.
• Gene expression profiles were analyzed using RNA sequencing.
• Changes in gene expression were correlated with enzyme treatment parameters (time, amount, temperature).

Main Results:

• Enzyme treatment significantly altered gene expression in a time-dependent manner.
• A subset of genes exhibited expression changes independent of enzyme treatment duration, amount, or temperature.
• Specific gene expression patterns were identified based on dissociation conditions.

Conclusions:

• Enzymatic dissociation introduces detectable gene expression alterations in scRNA-seq data.
• Understanding these enzyme-induced changes is critical for accurate data interpretation.
• This research provides essential data for selecting appropriate genes for analysis and refining cell isolation protocols.