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

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

Author information

1
Stephen A. Wynn, Institute for Vision Research, Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, Iowa.
2
Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, Iowa.

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.

KEYWORDS:

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

PMID:
28806854
PMCID:
PMC5609845
DOI:
10.1002/cpsc.30
[Indexed for MEDLINE]
Free PMC Article

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