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Derivation of Sendai-Virus-Reprogrammed Human iPSCs-Neuronal Precursors: In Vitro and In Vivo Post-grafting Safety Characterization
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SYSNO ASEP 0572487 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Derivation of Sendai-Virus-Reprogrammed Human iPSCs-Neuronal Precursors: In Vitro and In Vivo Post-grafting Safety Characterization Author(s) Shigyo, M. (US)
Kobayashi, Y. (JP)
Platoshyn, O. (US)
Maršala, S. (US)
Kato, T. (JP)
Takamura, N. (JP)
Yoshida, K. (JP)
Kishino, A. (JP)
Bravo-Hernandez, M. (US)
Juhás, Štefan (UZFG-Y) RID, ORCID
Juhásová, Jana (UZFG-Y) RID, ORCID
Studenovská, Hana (UMCH-V) RID, ORCID
Proks, Vladimír (UMCH-V) RID, ORCID
Ciacci, J. D. (US)
Maršala, M. (US)Article number 09636897231163232 Source Title Cell Transplantation. - : Sage - ISSN 0963-6897
Roč. 32, č. 1 (2023)Number of pages 15 s. Publication form Online - E Language eng - English Country GB - United Kingdom Keywords human-induced pluripotent stem cells ; neural precursor cells ; manual selection Subject RIV EB - Genetics ; Molecular Biology OECD category Cell biology Subject RIV - cooperation Institute of Macromolecular Chemistry - Macromolecular Chemistry Method of publishing Open access Institutional support UZFG-Y - RVO:67985904 ; UMCH-V - RVO:61389013 UT WOS 000955641300001 EID SCOPUS 85150860571 DOI 10.1177/09636897231163232 Annotation 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. Workplace Institute of Animal Physiology and Genetics Contact Jana Zásmětová, knihovna@iapg.cas.cz, Tel.: 315 639 554 Year of Publishing 2024 Electronic address https://journals.sagepub.com/doi/10.1177/09636897231163232
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