Medical College of Wisconsin
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From skin biopsy to neurons through a pluripotent intermediate under Good Manufacturing Practice protocols. Stem Cells Transl Med 2012 Jan;1(1):36-43

Date

12/01/2012

Pubmed ID

23197638

Pubmed Central ID

PMC3727693

DOI

10.5966/sctm.2011-0001

Abstract

The clinical application of human-induced pluripotent stem cells (hiPSCs) requires not only the production of Good Manufacturing Practice-grade (GMP-grade) hiPSCs but also the derivation of specified cell types for transplantation under GMP conditions. Previous reports have suggested that hiPSCs can be produced in the absence of animal-derived reagents (xenobiotics) to ease the transition to production under GMP standards. However, to facilitate the use of hiPSCs in cell-based therapeutics, their progeny should be produced not only in the absence of xenobiotics but also under GMP conditions requiring extensive standardization of protocols, documentation, and reproducibility of methods and product. Here, we present a successful framework to produce GMP-grade derivatives of hiPSCs that are free of xenobiotic exposure from the collection of patient fibroblasts, through reprogramming, maintenance of hiPSCs, identification of reprogramming vector integration sites (nrLAM-PCR), and finally specification and terminal differentiation of clinically relevant cells. Furthermore, we developed a primary set of Standard Operating Procedures for the GMP-grade derivation and differentiation of these cells as a resource to facilitate widespread adoption of these practices.

Author List

Karumbayaram S, Lee P, Azghadi SF, Cooper AR, Patterson M, Kohn DB, Pyle A, Clark A, Byrne J, Zack JA, Plath K, Lowry WE

Author

Michaela Patterson PhD Assistant Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Biopsy
Biotechnology
Cell Culture Techniques
Cell Separation
Cells, Cultured
Cellular Reprogramming
Fibroblasts
Gene Expression Regulation, Developmental
Guidelines as Topic
Humans
Laboratories
Male
Mice
Mice, SCID
Neural Stem Cells
Neurogenesis
Neurons
Pluripotent Stem Cells
Polymerase Chain Reaction
Quality Control
Reproducibility of Results
Skin