Good Manufacturing Practice–compliant human induced pluripotent stem cells: from bench to putative clinical products

• DOI: 10.1016/j.jcyt.2024.02.021
• Publication Date: 2024-03-05T08:20:12Z
• Authors: Juan J. Novoa; Inge M. Westra; Esther Steeneveld; Natascha Fonseca ...; Christiaan H. Are...; Bahareh Rajaei; Esmée Grundeken; Mehmet Yildiz; Wouter van der Valk; Alison Salvador; Françoise Carlotti; Pascale F. Dijkers; Heiko Locher; Cathelijne W. van...; Karine I. Raymond; Agnete Kirkeby; Christine L. Mummery; Ton J. Rabelink; Christian Freund; Pauline Meij; Brigitte Wieles
• Keywords: Germ layer; Regenerative Medicine; Good manufacturing practice

Background aimsFew human induced pluripotent stem cell (hiPSC) lines are Good Manufacturing Practice (GMP)-compliant, limiting the clinical use of hiPSC-derived products. Here, we addressed this by establishing and validating an in-house platform to produce GMP-compliant hiPSCs that would be appropriate for producing both allogeneic autologous products.MethodsOur standard research protocol production was adapted translated into a platform. In addition generation hiPSC, entails methodology donor recruitment, consent screening, material procurement, manufacture, in-process control, specific QC test validation, testing, product release, storage stability testing. For one run three runs were performed. Highest-quality selected establish master banks (MCBs).ResultsTwo MCBs successfully released under GMP conditions. They demonstrated safety (sterility, negative mycoplasma, endotoxins <5.0 EU/mL adventitious agents), identity (>75% cells expressing markers undifferentiated state, identical STR profile, normal karyotype in >20 metaphases), purity (negative residual vectors no plasmid integration genome) potency (expression at least two each germ layers). addition, directed differentiation somitoids (skeletal muscle precursors) six potential products from all layers achieved: pancreatic islets (endoderm), kidney organoids cardiomyocytes (mesoderm), keratinocytes, GABAergic interneurons inner-ear (ectoderm).ConclusionsWe developed validated generating hiPSC lines. The shown differentiate relevant our own other regenerative medicine interests.