Generation of Xeno-Free, cGMP-Compliant Patient-Specific iPSCs from Skin Biopsy

Curr Protoc Stem Cell Biol. 2017 Aug 14;42:4A.12.1-4A.12.14. doi: 10.1002/cpsc.30.

Abstract

This unit describes protocols for the generation of clinical-grade patient-specific induced pluripotent stem cell (iPSC)-derived retinal cells from patients with inherited retinal degenerative blindness. Specifically, we describe how, using xeno-free reagents in an ISO class 5 environment, one can isolate and culture dermal fibroblasts, generate iPSCs, and derive autologous retinal cells via 3-D differentiation. The universal methods described herein for the isolation of dermal fibroblasts and generation of iPSCs can be employed regardless of disease, tissue, or cell type of interest. © 2017 by John Wiley & Sons, Inc.

Keywords: cGMP; current Good Manufacturing Practice; fibroblasts; induced pluripotent stem cells; photoreceptor precursor cells; retina; xeno-free.

MeSH terms

  • Biopsy
  • Cell Culture Techniques / methods*
  • Cellular Reprogramming Techniques / methods*
  • Dermis* / metabolism
  • Dermis* / pathology
  • Fibroblasts* / metabolism
  • Fibroblasts* / pathology
  • Humans
  • Induced Pluripotent Stem Cells* / metabolism
  • Induced Pluripotent Stem Cells* / pathology
  • Retina* / metabolism
  • Retina* / pathology