Skip to content
Back to browse
RETINA Publication-derived

Retinal Bioprinted and Engineered Tissues for Regenerative Medicine

Full SOP with Academic or a plan

Source Maqueda et al., 2021 · Institut d'Investigació Biomèdica de Bellvitge (IDIBELL), Barcelona, Spain · 10.1016/j.biomaterials.2021.121049

Authors: Maria Maqueda, Jose Luis Mosquera, José García-Arumí, Anna Veiga, Anna Duarri Duration: 140 days 5 phases Cell source: Human iPSC

Abstract

This protocol describes the decellularization of mouse and porcine retinas to create acellular retinal ECM (arECM) scaffolds, followed by repopulation with hiPSC-derived retinal pigment epithelial (RPE) cells or ocular progenitor cells (OPCs). The protocol demonstrates that arECM scaffolds support cell engraftment, proliferation, differentiation, and organization into cell-type-specific clusters, providing a tissue-engineered model for studying retinal development and regeneration.

Cell source
Human iPSC
Application
Tissue engineering and regenerative medicine using acellular ECM scaffolds

Protocol overview

23 steps across 5 phases

Phase 1 · Retinal Isolation and Decellularization Day 0
  1. a Tissue collection and preparation
  2. b Retinal dissection
  3. c DNase I treatment
  4. d Non-ionic detergent treatment
  5. e Ionic detergent treatment
  6. f Washing with distilled water
  7. g Final PBS wash and storage
Phase 2 · Characterization of Acellular Retinal ECM Day 0-1
  1. a DNA quantification
  2. b Scanning electron microscopy (SEM) preparation
  3. c Proteomic analysis by LC-MS/MS
Phase 3 · hiPSC Culture and Differentiation Day 0-30
  1. a hiPSC maintenance culture
  2. b Ocular progenitor cell (OPC) differentiation
  3. c RPE differentiation and GFP transduction
Phase 4 · Recellularization of Acellular Retinal ECM Day 0-140
  1. a ECM equilibration
  2. b RPE cell preparation and seeding
  3. c OPC cell preparation and seeding
  4. d Medium replacement - day 1
  5. e Removal of non-attached cells - day 2
  6. f Continued culture and medium changes
Phase 5 · Characterization of Recellularized ECM Day 30-140
  1. a Histology and H&E staining
  2. b Immunohistochemistry (IHC)
  3. c Real-time quantitative PCR (qPCR)
  4. d Photoreceptor outer segment (POS) phagocytosis assay

Full SOP

Full SOPs need an account and a plan

Create an account, then get free Academic access (for verified academics) or choose a plan. Plans include step-by-step procedures, reagent concentrations, QC checklists and batch record templates.

Create an account

Already registered? Log in

Attribution

This SOP was written by Organthis based on the published method in Maqueda et al., 2021. Once the originating laboratory has confirmed it, the SOP is marked Verified.

Something wrong with this entry? Report an issue with this protocol

Need a commercial licence?
Use this protocol in your therapeutic or diagnostic pipeline.
Request Commercial Licence