Use cases
Epidermal organoids / ALI—not a plain HaCaT monolayer and not the default 130-day PSC hair-skin course.
| Use | Fit | Readout | Notes |
|---|---|---|---|
| Primary epidermal organoid expansion + infection / barrier studies | Recommended | Stratification markers, barrier function, infection readouts[1] | Serum-free recipes cut lot noise; log ALI vs expansion phases separately. |
| PSC hair-bearing skin organoids (long cycle) | Contrast | Hair follicles / appendages[2] | Cycles can run tens to >100 days; bank separately from primary expansion. |
| HaCaT monolayers as skin organoids | Do not mix | 2D confluence | 2D keratinocyte lines are not 3D epidermal organoids. |
In one line
Epidermal cells → serum-free niche expansion → optional ALI stratification → CK/barrier readout; PSC hair route as long-cycle contrast.
Protocol overview
- Ethics + cold chain; log site (foreskin / trunk…) and donor metadata
- Isolate epidermal cells; titrate collagenase / Dispase to clusters or primary keratinocytes
- Expand in serum-free epidermal niche (EGF, FGF, Noggin, R-spondin, A83-01…)[[1]]
- Brief Y-27632 at seeding; log passage and matrix / insert lot
- Optional: switch to air–liquid interface (ALI) for stratification and barrier readouts
- EpCAM / CK / E-Cadherin; mycoplasma; infection or drug arms with controls
- PSC hair-bearing long-cycle routes are contrast only[[2]]—do not mix with this expansion SOP