Preclinical Analysis of Fetal Human Mesencephalic Neural Progenitor Cell Lines: Characterization and Safety In Vitro and In Vivo

• Published in: Stem cells translational medicine 2016-09
• Main Authors: Moon, Jisook, Schwarz, Sigrid C, Lee, Hyun-Seob, Kang, Jun Mo, Lee, Young-Eun, Kim, Bona, Sung, Mi-Young, Höglinger, Günter, Wegner, Florian, Kim, Jin Su, Chung, Hyung-Min, Chang, Sung Woon, Cha, Kwang Yul, Kim, Kwang-Soo, Schwarz, Johannes
• Format: Article
• Language: English
• ISSN: 2157-6564

: We have developed a good manufacturing practice for long-term cultivation of fetal human midbrain-derived neural progenitor cells. The generation of human dopaminergic neurons may serve as a tool of either restorative cell therapies or cellular models, particularly as a reference for phenotyping region-specific human neural stem cell lines such as human embryonic stem cells and human inducible pluripotent stem cells. We cultivated 3 different midbrain neural progenitor lines at 10, 12, and 14 weeks of gestation for more than a year and characterized them in great detail, as well as in comparison with Lund mesencephalic cells. The whole cultivation process of tissue preparation, cultivation, and cryopreservation was developed using strict serum-free conditions and standardized operating protocols under clean-room conditions. Long-term-cultivated midbrain-derived neural progenitor cells retained stemness, midbrain fate specificity, and floorplate markers. The potential to differentiate into authentic A9-specific dopaminergic neurons was markedly elevated after prolonged expansion, resulting in large quantities of functional dopaminergic neurons without genetic modification. In restorative cell therapeutic approaches, midbrain-derived neural progenitor cells reversed impaired motor function in rodents, survived well, and did not exhibit tumor formation in immunodeficient nude mice in the short or long term (8 and 30 weeks, respectively). We conclude that midbrain-derived neural progenitor cells are a promising source for human dopaminergic neurons and suitable for long-term expansion under good manufacturing practice, thus opening the avenue for restorative clinical applications or robust cellular models such as high-content or high-throughput screening. For many years, human fetal midbrain-derived neural stem or progenitor cells have been considered a potential source for restorative therapy in patients with Parkinson's disease. However, there have been considerable concerns with respect to limited expansion, early senescence, and loss of pluripotency. This study investigated such cells thoroughly and developed techniques that allow for long-term stable expansion, overcoming the above concerns. The cells were found to be safe and potent, allowing clinical application.