title: Preclinical Analysis of Fetal Human Mesencephalic Neural Progenitor Cell Lines: Characterization and Safety In Vitro and In Vivo.
abstract: 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. Stem Cells Translational Medicine 2017;6:576-588.
authors.affiliation: Department of Biotechnology, College of Life Science, CHA University, Seongnam-si, Gyeonggi-do, Korea.|General Research Division, Korea Research-Driven Hospital, Bundang CHA Medical Center, CHA University, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Jisook
authors.initials: J
authors.lastname: Moon
authors.affiliation: German Center for Neurodegenerative Diseases, Technical University Munich, Munich, Germany.
authors.forename: Sigrid C
authors.initials: SC
authors.lastname: Schwarz
authors.affiliation: General Research Division, Korea Research-Driven Hospital, Bundang CHA Medical Center, CHA University, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Hyun-Seob
authors.initials: HS
authors.lastname: Lee
authors.affiliation: General Research Division, Korea Research-Driven Hospital, Bundang CHA Medical Center, CHA University, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Jun Mo
authors.initials: JM
authors.lastname: Kang
authors.affiliation: General Research Division, Korea Research-Driven Hospital, Bundang CHA Medical Center, CHA University, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Young-Eun
authors.initials: YE
authors.lastname: Lee
authors.affiliation: Development Division, CHA Biotech, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Bona
authors.initials: B
authors.lastname: Kim
authors.affiliation: Development Division, CHA Biotech, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Mi-Young
authors.initials: MY
authors.lastname: Sung
authors.affiliation: German Center for Neurodegenerative Diseases, Technical University Munich, Munich, Germany.
authors.forename: Günter
authors.initials: G
authors.lastname: Höglinger
authors.affiliation: Department of Neurology, Hannover Medical School, Hannover, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Wegner
authors.affiliation: Molecular Imaging Research Center, Korea Institute of Radiological and Medical Sciences, Seoul, Korea.
authors.forename: Jin Su
authors.initials: JS
authors.lastname: Kim
authors.affiliation: Department of Stem Cell Biology, Graduate School of Medicine, Konkuk University, Gwangjin-gu, Seoul, Korea.
authors.forename: Hyung-Min
authors.initials: HM
authors.lastname: Chung
authors.affiliation: Department of Obstetrics and Gynecology, CHA Bundang Medical Center, CHA University, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Sung Woon
authors.initials: SW
authors.lastname: Chang
authors.affiliation: General Research Division, Korea Research-Driven Hospital, Bundang CHA Medical Center, CHA University, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Kwang Yul
authors.initials: KY
authors.lastname: Cha
authors.affiliation: Molecular Neurobiology Laboratory, Department of Psychiatry, Program in Neuroscience and Harvard Stem Cell Institute, McLean Hospital/Harvard Medical School, Belmont, Massachusetts, USA.
authors.forename: Kwang-Soo
authors.initials: KS
authors.lastname: Kim
authors.affiliation: German Center for Neurodegenerative Diseases, Technical University Munich, Munich, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Schwarz
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques; D002460:Cell Line; D049109:Cell Proliferation; D004195:Disease Models, Animal; D059290:Dopaminergic Neurons / Q000378:metabolism / Q000502:physiology*; D005260:Female; D005865:Gestational Age; D006801:Humans; D008297:Male; D008636:Mesencephalon / Q000196:embryology*; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009043:Motor Activity; D058953:Neural Stem Cells / Q000378:metabolism / Q000502:physiology*; D055495:Neurogenesis; D016627:Oxidopamine; D020734:Parkinsonian Disorders / Q000139:chemically induced / Q000473:pathology / Q000503:physiopathology / Q000601:surgery*; D010641:Phenotype; D017207:Rats, Sprague-Dawley; D020127:Recovery of Function; D018570:Risk Assessment; D033581:Stem Cell Transplantation / Q000009:adverse effects / Q000379:methods*; D013724:Teratoma / Q000209:etiology / Q000473:pathology; D013997:Time Factors
chemical_list: D015415:Biomarkers; D016627:Oxidopamine
keywords: Dopaminergic differentiation; Human fetal midbrain tissue; Neural progenitor cells; Parkinson's disease; Preclinical safety; Rodents; Transplantation