Page 2 - R. Organoid

[R. Organoid-1]

Characterization of iPSC-derived midbrain organoids from

Parkinson's disease patient

Yunsu Bang¹˙#˙*, Juhyun Choi¹˙#, Ki Soon Kim¹, Jong Gu Lee¹, Woo Yong Oh¹, Yoonsook Lee¹˙*

¹Clinical Research Division, NIFDS, MFDS, Cheongju 28159, Korea

Parkinson's disease(PD) is the one of common neurodegenerative diseases, that results from the loss of
neuromelanin(NM)-containing dopaminergic(DA) neurons. To overcome the limitations of animal degenerative
models for human phenotypes, human induced Pluripotent Stem Cell(iPSCs) are being widely used nowdays. In this

study, we investigated the characterization of midbrain organoids(MOs) whether these can be the proper model for

research and drug screening on neurodegenerative diseases.

At first, we differentiated MOs from human iPSCs by 3D culture system. And then we checked the identified markers
for representing DA neurons on culture day 0, 24, 44, 64, 84 by real time PCR, WB, and IHC. The differentiated MOs

showed that DA progenitors(FoxA2, LMX1A and LMX1B), neuronal microtubule(Tuj1, MAP2) and DA neuron(TH,
GIRK2) markers including accumulation of alpha-synuclein. First of all, the NM granules were observed in organoids,

not shown in 2D-cultured iPSCs. NM granules of patient's MOs were detected lower than those of normals', that
shows the major characteristic of PD.

Our study showed that the MOs had several features of the human midbrain. Even though more assessments remain
to be elucidated, we propose that iPSCs-derived organoids are useful tool for researches.