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SKIN Publication-derived

Skin Organ-on-Chip Models

Full SOP with Academic or a plan

Source Quílez et al. · Columbia University Irving Medical Center, New York, USA · 10.1002/adhm.202400405

Authors: Quílez C, Jeon EY, Pappalardo A, Pathak P, Abaci HE Duration: 32 days 9 phases Cell source: Human iPSC

Abstract

Novel approach generating skin organoids in a 3D bioprinted spindle-shaped hydrogel device with defined ECM (Collagen I + Laminin 511). Overcomes the inverted anatomy limitation of conventional skin organoids and achieves 2-fold keratinocyte and 8-fold hair follicle increase compared to free-floating conditions.

Cell source
Human iPSC
Application
Organ-on-Chip Skin Models

Protocol overview

31 steps across 9 phases

Phase 1 · mESC Maintenance Ongoing maintenance prior to day 0
  1. a Maintain mESCs in feeder-free conditions
Phase 2 · Initial Embryoid Body (EB) Formation Day 0-Day 3
  1. a Detach and resuspend mESC colonies
  2. b Seed cells in ultra-low attachment plate for EB formation
  3. c Culture EBs in incubator
  4. d Medium supplementation on Day 1
Phase 3 · ECM Hydrogel Encapsulation (Day 3) Day 3
  1. a Prepare collagen I hydrogel solution
  2. b Embed EBs in hydrogel
  3. c Crosslink hydrogel
  4. d Add differentiation medium with morphogens
Phase 4 · Early Differentiation (Day 4 onwards) Day 4-Day 8
  1. a Add neural crest differentiation factors on Day 4
  2. b Transfer to maturation culture on Day 8
Phase 5 · Maturation Phase (Day 8-Day 32) Day 8-Day 32
  1. a Maintain maturation culture with regular medium changes
  2. b Monitor organoid growth and morphology
Phase 6 · 3D Bioprinting of Spindle Device (Optional—for enhanced growth) Device fabrication prior to Day 3 seeding
  1. a Design and 3D bioprint spindle device
  2. b Clean and sterilize printed devices
  3. c Prepare devices for cell seeding
Phase 7 · PSO Generation in Spindle Device (Alternative to standard plate culture) Day 3-Day 32
  1. a Seed EBs in spindle device on Day 3
  2. b Culture spindle devices with initial differentiation medium
  3. c Add neural crest factors on Day 4
  4. d Transition to maturation medium on Day 8
  5. e Continue maturation with regular medium changes
Phase 8 · Microfluidic Device Fabrication and Setup (Optional—for gradient control) Device fabrication prior to Day 2 seeding
  1. a 3D bioprint microfluidic device with gradient channels
  2. b 3D print perfusion chamber in PLA
  3. c Assemble microfluidic and perfusion system
  4. d Prepare devices for cell seeding
Phase 9 · PSO Generation in Microfluidic Device with Morphogen Gradients Day 2-Day 32
  1. a Seed EBs in microfluidic device on Day 2
  2. b Culture EBs with E6 medium in lateral chambers
  3. c Establish epidermal differentiation gradient on Day 4
  4. d Establish dermal differentiation gradient on Day 5
  5. e Transition to maturation medium on Day 10
  6. f Maintain maturation culture through Day 32

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Attribution

This SOP was written by Organthis based on the published method in Quílez et al.. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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