Validating human induced pluripotent stem cell-specific quality control tests for the release of an intermediate drug product in a Good Manufacturing Practice quality system
Cytotherapy, vol. 26, pp. 1105–1117
Abstract
One of the challenges in Good Manufacturing Practice (GMP)-compliant human induced pluripotent stem cell (hiPSC) production is the validation of quality control (QC) tests specific for hiPSCs, which are required for GMP batch release. This study presents a comprehensive description of the validation process for hiPSC-specific GMP-compliant QC assays; more specifically, the validation of assays to assess the potential presence of residual episomal vectors (REVs), the expression of markers of the undifferentiated state and the directed differentiation potential of hiPSCs. Critical aspects and specific acceptance criteria were formulated in a validation plan prior to assay validation. Assay specificity, sensitivity and reproducibility were tested, and the equipment used for each assay was subjected to performance qualification. A minimum input of 20 000 cells (120 ng of genomic DNA) was defined for accurate determination of the presence of REVs. Furthermore, since vector loss in hiPSC lines is a passage-dependent process, we advocate screening for REVs between passages eight and 10, as testing at earlier passages might lead to unnecessary rejection of hiPSC lines. The cutoff value for assessment of markers of the undifferentiated state was set to the expression of at least three individual markers on at least 75% of the cells. When multi-color flow cytometry panels are used, a fluorescence minus one control is advised to ensure the control for fluorescent spread. For the assay to assess the directed differentiation potential, the detection limit was set to two of three positive lineage-specific markers for each of the three individual germ layers. All of our assays proved to be reproducible and specific. Our data demonstrate that our implemented analytical procedures are suitable as QC assays for the batch release of GMP-compliant hiPSCs.
Authors 12
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Leiden University Medical Center
Affiliation as printed
Department of Internal Medicine, Leiden University Medical Center, Leiden, the Netherlands
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Leiden University Medical Center
Affiliation as printed
Center for Cell and Gene Therapy, Leiden University Medical Center, Leiden, the Netherlands
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Leiden University Medical Center
Affiliation as printed
Department of Internal Medicine, Leiden University Medical Center, Leiden, the Netherlands
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Leiden University Medical Center
Affiliation as printed
Center for Cell and Gene Therapy, Leiden University Medical Center, Leiden, the Netherlands
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Leiden University Medical Center
Affiliation as printed
Department of Internal Medicine, Leiden University Medical Center, Leiden, the Netherlands
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Albert Blanch-Asensio Aachen Department of Anatomy and Embryology Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)
Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine
Affiliation as printed
Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, the Netherlands; Novo Nordisk Foundation Center for Stem Cell Medicine, Leiden University Medical Center, the Netherlands
Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, The Netherlands
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Richard P. Davis Aachen Department of Anatomy and Embryology Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)
Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine
Affiliation as printed
Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, the Netherlands; Novo Nordisk Foundation Center for Stem Cell Medicine, Leiden University Medical Center, the Netherlands
Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, The Netherlands
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Leiden University Medical Center
Affiliation as printed
Department of Internal Medicine, Leiden University Medical Center, Leiden, the Netherlands
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Leiden University Medical Center
Affiliation as printed
Leiden University Medical Center hiPSC Hotel, Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, the Netherlands
LUMC hiPSC Hotel, Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, the Netherlands
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Ton J. Rabelink corresponding Aachen Department of Internal Medicine Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW)
Leiden University Medical Center · Novo Nordisk Foundation Center for Stem Cell Medicine
Affiliation as printed
Department of Internal Medicine, Leiden University Medical Center, Leiden, the Netherlands; Novo Nordisk Foundation Center for Stem Cell Medicine, Leiden University Medical Center, the Netherlands. Electronic address: a.j.rabelink@lumc.nl
Department of Internal Medicine, Leiden University Medical Center, Leiden, the Netherlands
The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, The Netherlands
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Leiden University Medical Center
Affiliation as printed
Center for Cell and Gene Therapy, Leiden University Medical Center, Leiden, the Netherlands
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Leiden University Medical Center
Affiliation as printed
Department of Internal Medicine, Leiden University Medical Center, Leiden, the Netherlands
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Cited by patents worldwide 1 (Lens.org)
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