Title

Human iPSC-Derived Assembloid of Sensory Pathway: A Platform for Investigating E/I Imbalance in Autism Spectrum Disorders

Alternative Title

중추 신경계 감각 경로 모델링: 자폐스펙트럼장애의 흥분성/억제성 신경망 불균형 연구를 위한 인간 유도만능줄기세포 유래 어셈블로이드

DGIST Authors

Juwon Lee ; Sung Bae Lee ; Jinsoo Seo

Advisor

이성배

Co-Advisor(s)

Jinsoo Seo

Issued Date

2025

Awarded Date

2025-08-01

Type

Thesis

Description

induced pluripotent stem cells (iPSCs), assembloid, thalamocortical circuit, microglia, autism spectrum disorder (ASD)

Table Of Contents

Ⅰ. Introduction 1
1.1 Brain Organoids as a Human-Based Model 1
1.2 Assembloid for Modeling Complex Brain Circuits 2
1.3 Autism Spectrum Disorder 3
1.3.1 Sensory abnormalities and ASD 4
1.3.2 ASD-related genes 5
1.4 Challenges in Studying ASD in Humans 7
IⅠ. Materials and Methods 11
2.1 Cell culture 11
2.1.1 Human induced pluripotent stem cells (hiPSCs) 11
2.1.2 Generation of ventral thalamic organoid (vThO), ganglionic eminence organoid (GeO), and cerebral cortex organoid (CxO) from hiPSCs 11
2.1.3 HPC Differentiation from iPSCs 13
2.1.4 Microglia Differentiation from HPC 14
2.1.5 CRISPR/Cas9-mediated gene editing of CNTNAP2 and SHANK3 15
2.2 Generation of assembloid 16
2.3 Cryosection and immunohistochemistry 16
2.4 Real-time qPCR 17
ⅠII. Results 18
3.1 Previous Modeling Strategies and Opportunities for Extension 18
3.2 Designing the structure of CS assembloid and modifying protocols 23
3.3 Optimization of culture conditions for ventral thalamic organoids 27
3.4 Establishing a CS assembloid by fusing ventral thalamic, ganglionic eminence, and cortical organoids 32
3.5 iPSC-derived microglia like cells in Cerebral cortex organoid 37
3.6 CRISPR/Cas9-based targeting for CNTNAP2 and SHANK3 in iPSCs 42
IV. Disccusion 46
V. References 49
Abstract in Korean 53

URI

https://scholar.dgist.ac.kr/handle/20.500.11750/59811
http://dgist.dcollection.net/common/orgView/200000889586

DOI

10.22677/THESIS.200000889586

Degree

Master

Department

Department of Brain Sciences

Publisher

DGIST

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