title: From skin biopsy to neurons through a pluripotent intermediate under Good Manufacturing Practice protocols.
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.
authors.affiliation: Eli and Edythe Broad Center for Regenerative Medicine.
authors.forename: Saravanan
authors.initials: S
authors.lastname: Karumbayaram
authors.forename: Peiyee
authors.initials: P
authors.lastname: Lee
authors.forename: Soheila F
authors.initials: SF
authors.lastname: Azghadi
authors.forename: Aaron R
authors.initials: AR
authors.lastname: Cooper
authors.forename: Michaela
authors.initials: M
authors.lastname: Patterson
authors.forename: Donald B
authors.initials: DB
authors.lastname: Kohn
authors.forename: April
authors.initials: A
authors.lastname: Pyle
authors.forename: Amander
authors.initials: A
authors.lastname: Clark
authors.forename: James
authors.initials: J
authors.lastname: Byrne
authors.forename: Jerome A
authors.initials: JA
authors.lastname: Zack
authors.forename: Kathrin
authors.initials: K
authors.lastname: Plath
authors.forename: William E
authors.initials: WE
authors.lastname: Lowry
mesh_terms: D000818:Animals; D001706:Biopsy / Q000592:standards; D001709:Biotechnology / Q000592:standards*; D018929:Cell Culture Techniques / Q000592:standards; D002469:Cell Separation / Q000592:standards; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D005347:Fibroblasts / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental; D017408:Guidelines as Topic; D006801:Humans; D007753:Laboratories / Q000592:standards*; D008297:Male; D051379:Mice; D016513:Mice, SCID; D058953:Neural Stem Cells / Q000502:physiology*; D055495:Neurogenesis; D009474:Neurons / Q000502:physiology*; D039904:Pluripotent Stem Cells / Q000502:physiology*; D016133:Polymerase Chain Reaction / Q000592:standards; D011786:Quality Control; D015203:Reproducibility of Results; D012867:Skin / Q000166:cytology*