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1
hiPSC Culture
Pluripotent cells expanded under defined conditions
Key point:
Quality control: karyotype, markers
Directed epidermal differentiation
Growth-factor protocol → keratinocyte lineage
Key point:
Marker panel: KRT14, p63
Timeframe: days–weeks
Epidermal organoid formation
3D self-assembled structures with stratification
Key point:
Histology: basal → spinous → granular layers
EV biogenesis & isolation
Collect conditioned medium → isolate EVs
Methods:
Ultracentrifugation / density gradient / SEC
Quality: NTA, TEM, marker panel (CD9/CD63)
Functional cargo & assays
Proteomics, miRNA, and in vitro tests
Assays:
Fibroblast migration, keratinocyte proliferation
Dose-response and blocking experiments
Skin regeneration outcomes
In vivo wound models & histological scoring
Readouts:
Re-epithelialization, vascularization, ECM remodelling
Safety: inflammation, off-target effects
Conceptual figure: Development of PSC-derived epidermal organoids → EVs → skin regeneration (6-step summary)
Figure legend (for Preview):
This schematic summarises the experimental pipeline described in the selected paper: (1) expansion of hiPSCs, (2) directed differentiation to epidermal progenitors, (3) formation of 3D epidermal organoids with stratified architecture, (4) isolation and characterisation of extracellular vesicles (EVs) from organoid conditioned medium, (5) biochemical and functional assays demonstrating EV cargo and regenerative effects in vitro, and (6) in vivo skin repair outcomes including re-epithelialisation and ECM remodelling. Each step lists critical quality controls and methods that should be briefly discussed in the Preview (e.g., markers, NTA/TEM, proteomics, functional blocking studies, histology).
Design notes: horizontal layout for single-figure Preview (fits journal style). Use this figure as the single required schematic (one figure). Keep figure file as vector (SVG/PDF) for submission to ensure clarity at publication size.
Conceptual figure: Development of PSC-derived epidermal organoids → EVs → skin regeneration (6-step summary)
Figure legend (for Preview): This schematic summarises the experimental pipeline described in the selected paper: (1) expansion of hiPSCs, (2) directed differentiation to epidermal progenitors, (3) formation of 3D epidermal organoids with stratified architecture, (4) isolation and characterisation of extracellular vesicles (EVs) from organoid conditioned medium, (5) biochemical and functional assays demonstrating EV cargo and regenerative effects in vitro, and (6) in vivo skin repair outcomes including re-epithelialisation and ECM remodelling. Each step lists critical quality controls and methods that should be briefly discussed in the Preview (e.g., markers, NTA/TEM, proteomics, functional blocking studies, histology). Design notes: horizontal layout for single-figure Preview (fits journal style). Use this figure as the single required schematic (one figure). Keep figure file as vector (SVG/PDF) for submission to ensure clarity at publication size.