Establishment of stable iPS-derived human neural stem cell lines suitable for cell therapies

Jessica Rosati, Daniela Ferrari, Filomena Altieri, Silvia Tardivo, Claudia Ricciolini, Caterina Fusilli, Cristina Zalfa, Daniela C. Profico, Francesca Pinos, Laura Bernardini, Barbara Torres, Isabella Manni, Giulia Piaggio, Elena Binda, Massimiliano Copetti, Giuseppe Lamorte, Tommaso Mazza, Massimo Carella, Maurizio Gelati, Enza Maria Valente, Antonio Simeone, Angelo L. Vescovi

Establishing specific cell lineages from human induced pluripotent stem cells (hiPSCs) is vital for cell therapy approaches in regenerative medicine, particularly for neurodegenerative disorders. While neural precursors have been induced from hiPSCs, the establishment of hiPSC-derived human neural stem cells (hiNSCs), with characteristics that match foetal hNSCs and abide by cGMP standards, thus allowing clinical applications, has not been described. We generated hiNSCs by a virus-free technique, whose properties recapitulate those of the clinical-grade hNSCs successfully used in an Amyotrophic Lateral Sclerosis (ALS) phase I clinical trial. Ex vivo, hiNSCs critically depend on exogenous mitogens for stable self-renewal and amplification and spontaneously differentiate into astrocytes, oligodendrocytes and neurons upon their removal. In the brain of immunodeficient mice, hiNSCs engraft and differentiate into neurons and glia, without tumour formation. These findings now warrant the establishment of clinical-grade, autologous and continuous hiNSC lines for clinical trials in neurological diseases such as Huntington's, Parkinson's and Alzheimer's, among others.