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Updated: Mar 17, 2026
A Two-Step Strategy that Combines Epigenetic Modification and Biomechanical Cues to Generate Mammalian Pluripotent Cells
Published on: August 29, 2020
Induced Pluripotent Stem Cell Differentiation and Three-Dimensional Tissue Formation Attenuate Clonal Epigenetic
Anastasia Petrova1,2,3, Antonio Capalbo4, Laureen Jacquet1
11 Assisted Conception Unit, Stem Cell Laboratory, Division of Women's Health, Women's Health Academic Centre, King's College London , London, United Kingdom .
Induced pluripotent stem cells (iPSCs) showed stable genomes and miRNAomes after culture. However, DNA methylation differences were observed, particularly in TCHH, which did not impact epidermal differentiation in 3D models.
Area of Science:
- Stem Cell Biology
- Epigenetics
- Tissue Engineering
Background:
- Epigenetic variations in pluripotent stem cells can affect their behavior.
- Existing research primarily uses standard monolayer cell cultures.
- Three-dimensional (3D) models offer a more accurate representation of in vivo conditions.
Purpose of the Study:
- To comparatively analyze two induced pluripotent stem cell (iPSC) lines for 3D human epidermal equivalent (HEE) engineering.
- To assess genomic stability, miRNAome, and DNA methylation profiles of iPSCs.
- To investigate the impact of epigenetic differences on epidermal differentiation.
Main Methods:
- Exome sequencing and array comparative genomic hybridization (CGH) for genomic stability.
- miRNA array and DNA methylation array for epigenetic profiling.
- Three-dimensional (3D) tissue engineering to create human epidermal equivalents (HEE).
- Immunostaining for protein expression analysis.
Main Results:
- Genomes and miRNAomes of iPSC lines remained stable after 3 months of feeder-free culture.
- Approximately 7% of CpG sites showed differential DNA methylation between the two iPSC lines.
- One iPSC clone (iKCL004) retained a somatic methylation signature at the TCHH gene, while another (iKCL011) matched embryonic stem cell methylation.
- Observed differences in TCHH methylation did not significantly affect its expression in 3D HEE.
Conclusions:
- Induced pluripotent stem cell (iPSC) lines exhibit genomic and miRNAome stability during culture.
- Significant DNA methylation differences exist between iPSC lines, particularly at the TCHH locus.
- Epidermal differentiation and 3D tissue formation may overcome variations in the iPSC methylome.
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