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

Juan J Novoa; Inge M Westra; Esther Steeneveld; Natascha Fonseca Neves; Christiaan H Arendzen; Bahareh Rajaei; Esmée Grundeken; Mehmet Yildiz; Wouter van der Valk; Alison Salvador; Françoise Carlotti; Pascale F Dijkers; Heiko Locher; Cathelijne W van den Berg; Karine I Raymond; Agnete Kirkeby; Christine L Mummery; Ton J Rabelink; Christian Freund; Pauline Meij; Brigitte Wieles

doi:10.1016/j.jcyt.2024.02.021

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

Cytotherapy. 2024 Mar 5:S1465-3249(24)00070-7. doi: 10.1016/j.jcyt.2024.02.021. Online ahead of print.

Authors

Juan J Novoa¹, Inge M Westra², Esther Steeneveld¹, Natascha Fonseca Neves³, Christiaan H Arendzen⁴, Bahareh Rajaei¹, Esmée Grundeken⁵, Mehmet Yildiz⁶, Wouter van der Valk⁷, Alison Salvador⁸, Françoise Carlotti¹, Pascale F Dijkers⁶, Heiko Locher⁷, Cathelijne W van den Berg⁹, Karine I Raymond⁶, Agnete Kirkeby⁸, Christine L Mummery⁶, Ton J Rabelink¹⁰, Christian Freund⁵, Pauline Meij², Brigitte Wieles¹

Affiliations

¹ Department of Internal Medicine, Leiden University Medical Center, Leiden, The Netherlands.
² Center for Cell and Gene Therapy, Leiden University Medical Center, Leiden, The Netherlands; The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, Leiden, The Netherlands.
³ Center for Cell and Gene Therapy, Leiden University Medical Center, Leiden, The Netherlands.
⁴ LUMC hiPSC Hotel, Department of Anatomy and Embryology, Leiden University Medical Center, Leiden, The Netherlands.
⁵ LUMC hiPSC Hotel, 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, Leiden, The Netherlands.
⁶ 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, Leiden, The Netherlands.
⁷ OtoBiology Leiden, Department of Otorhinolaryngology and Head & Neck Surgery, Leiden University Medical Center, Leiden, The Netherlands; The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center, Leiden, The Netherlands.
⁸ Department of Neuroscience, University of Copenhagen, Copenhagen, Denmark; The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), University of Copenhagen, Copenhagen, Denmark.
⁹ 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, Leiden, The Netherlands.
¹⁰ 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, Leiden, The Netherlands. Electronic address: a.j.rabelink@lumc.nl.

PMID: 38483359
DOI: 10.1016/j.jcyt.2024.02.021

Abstract

Background aims: Few 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 and autologous hiPSC-derived products.

Methods: Our standard research protocol for hiPSCs production was adapted and translated into a GMP-compliant platform. In addition to the generation of GMP-compliant hiPSC, the platform entails the methodology for donor recruitment, consent and screening, donor material procurement, hiPSCs manufacture, in-process control, specific QC test validation, QC testing, product release, hiPSCs storage and stability testing. For platform validation, one test run and three production runs were performed. Highest-quality lines were selected to establish master cell banks (MCBs).

Results: Two MCBs were successfully released under GMP conditions. They demonstrated safety (sterility, negative mycoplasma, endotoxins <5.0 EU/mL and negative adventitious agents), cell identity (>75% of cells expressing markers of undifferentiated state, identical STR profile, normal karyotype in >20 metaphases), purity (negative residual vectors and no plasmid integration in the genome) and potency (expression of at least two of the three markers for each of the three germ layers). In addition, directed differentiation to somitoids (skeletal muscle precursors) and six potential clinical products from all three germ layers was achieved: pancreatic islets (endoderm), kidney organoids and cardiomyocytes (mesoderm), and keratinocytes, GABAergic interneurons and inner-ear organoids (ectoderm).

Conclusions: We successfully developed and validated a platform for generating GMP-compliant hiPSC lines. The two MCBs released were shown to differentiate into clinical products relevant for our own and other regenerative medicine interests.

Keywords: GMP production; cell therapy; hiPSC-derived products; human induced pluripotent stem cells; quality control testing.