Derivation of Sendai-Virus-Reprogrammed Human iPSCs-Neuronal Precursors: In Vitro and In Vivo Post-grafting Safety Characterization

0572487 - ÚŽFG 2024 RIV GB eng J - Journal Article
Shigyo, M. - Kobayashi, Y. - Platoshyn, O. - Maršala, S. - Kato, T. - Takamura, N. - Yoshida, K. - Kishino, A. - Bravo-Hernandez, M. - Juhás, Štefan - Juhásová, Jana - Studenovská, Hana - Proks, Vladimír - Ciacci, J. D. - Maršala, M.
Derivation of Sendai-Virus-Reprogrammed Human iPSCs-Neuronal Precursors: In Vitro and In Vivo Post-grafting Safety Characterization.
Cell Transplantation. Roč. 32, č. 1 (2023), č. článku 09636897231163232. ISSN 0963-6897. E-ISSN 1555-3892
Institutional support: RVO:67985904 ; RVO:61389013
Keywords : human-induced pluripotent stem cells * neural precursor cells * manual selection
OECD category: Cell biology; Polymer science (UMCH-V)
Impact factor: 3.3, year: 2022
Method of publishing: Open access
https://journals.sagepub.com/doi/10.1177/09636897231163232

The critical requirements in developing clinical-grade human-induced pluripotent stem cells-derived neural precursors (hiPSCs-NPCs) are defined by expandability, genetic stability, predictable in vivo post-grafting differentiation, and acceptable safety profile. Here, we report on the use of manual-selection protocol for generating expandable and stable human NPCs from induced pluripotent stem cells. The hiPSCs were generated by the reprogramming of peripheral blood mononuclear cells with Sendai-virus (SeV) vector encoding Yamanaka factors. After induction of neural rosettes, morphologically defined NPC colonies were manually harvested, re-plated, and expanded for up to 20 passages. Established NPCs showed normal karyotype, expression of typical NPCs markers at the proliferative stage, and ability to generate functional, calcium oscillating GABAergic or glutamatergic neurons after in vitro differentiation. Grafted NPCs into the striatum or spinal cord of immunodeficient rats showed progressive maturation and expression of early and late human-specific neuronal and glial markers at 2 or 6 months post-grafting. No tumor formation was seen in NPCs-grafted brain or spinal cord samples. These data demonstrate the effective use of in vitro manual-selection protocol to generate safe and expandable NPCs from hiPSCs cells. This protocol has the potential to be used to generate GMP (Good Manufacturing Practice)-grade NPCs from hiPSCs for future clinical use.
Permanent Link: https://hdl.handle.net/11104/0343144