Session Information
Date: Tuesday, June 21, 2016
Session Title: Parkinson's disease: Pathophysiology
Session Time: 12:30pm-2:00pm
Location: Exhibit Hall located in Hall B, Level 2
Objective: To generate and characterize human pluripotent stem cell (iPSCs) – derived midbrain cerebral organoids of patients with Parkinson’s disease.
Background: Parkinson’s disease is a neurodegenerative disorder characterized by a progressive loss of midbrain dopaminergic neurons. Several genetic causes have been identified in familial and sporadic cases. However an additional effort is required to understand the molecular cascade that lead to neuronal alpha-synuclein accumulation and the neurodegenerative process. In this project, we exploit the potential of human stem cell-derived three-dimensional organoids to address these challenges.
Methods: Fibroblasts obtained from skin biopsies of one patient and two controls were reprogrammed to iPSCs using a CytoTune-iPS 2.0 Sendai Reprogramming Kit (Life Technologies) based on the viral transduction of the factors Oct, Sox2, Klf4 and c-Myc. Cell lines were evaluated for the expression of the stemness markers SSEA-4, TRA1-60, Oct, Sox 2 through ICC. Karyotypes were evaluated to exclude genetic abnormalities. We differentiated iPSC lines into organoids using a modification of a method by Lancaster et al., 2013 and Tieng et al. 2014. After 40 days of culture in spinner flasks, organoids were collected and analyzed by ICH and WB. CGase chemiluminescent enzymatic assay was performed using 4MUβG 10 mM as substrate and NaTaurocholate 50 mM incubated at 37°C for 30 min.
Results: Organoids from iPSCs of a PD patient harbouring a heterozygous L444P GBA, and two controls contained neurons diffusely positive for staining with TH and GIRK2, showing thus a nigral dopaminergic identity. Phosphorylated-alpha-synuclein appeared to be expressed in patients’ iPSC-derived neurons and in controls. GCase enzyme showed a significant decreased activity in patient organoids.
Conclusions: Midbrain organoids may represent a suitable tool to dissect multiple aspects of Parkinson’s disease pathogenesis.
To cite this abstract in AMA style:
E. Frattini, G. Monzio Compagnoni, S. Salani, P. Rinchetti, M. Nizzardo, M. Baccarin, N. Bresolin, G.P. Comi, S.P. Corti, A. Di Fonzo. Generation and characterization of midbrain organoids from iPSCs of familial Parkinson’s disease [abstract]. Mov Disord. 2016; 31 (suppl 2). https://www.mdsabstracts.org/abstract/generation-and-characterization-of-midbrain-organoids-from-ipscs-of-familial-parkinsons-disease/. Accessed October 16, 2025.« Back to 2016 International Congress
MDS Abstracts - https://www.mdsabstracts.org/abstract/generation-and-characterization-of-midbrain-organoids-from-ipscs-of-familial-parkinsons-disease/