Development of Fabry disease kidney organoid-on-a-chip for gradient drug screening
- 주제어 (키워드) Kidney organoids , Fabry disease , Microfluidic gradient chip , Enzyme replacement therapy (ERT) , Drug screening
- 발행기관 서강대학교 일반대학원
- 지도교수 정봉근
- 발행년도 2024
- 학위수여년월 2024. 8
- 학위명 석사
- 학과 및 전공 일반대학원 융합의생명공학과협동과정
- 실제 URI http://www.dcollection.net/handler/sogang/000000079115
- UCI I804:11029-000000079115
- 본문언어 영어
- 저작권 서강대학교 논문은 저작권 보호를 받습니다.
초록 (요약문)
The recent development of the kidney organoids provides a biochemical platform for investigating disease mechanism and drug screening. These three-dimensional (3D) constructs summarize key aspects of the human kidney, aiding in the study of complex disease processes. However, the conventional in vitro models fail to fully reflect the complexity of native tissues. To address this issue, there is a need to develop the novel culture systems that support physiologically improved kidney organoid differentiation. In this study, I investigate the efficacy of a microfluidic gradient chip system in enhancing the differentiation, maturation, and functionality of normal and Fabry disease kidney organoids. The results demonstrate that the fluidic flow promotes superior vascularization, nutrient uptake, and gene expression associated with kidney maturation as compared to static culture conditions. Moreover, the increase in expression of specific markers and upregulation of kidney-related genes in microfluidic chip systems effectively enhance the differentiation, maturation, and functional potential of both normal and Fabry disease kidney organoids. Furthermore, the microfluidic gradient chip systems demonstrate improved sensitivity in detecting therapeutic effects, such as the restoration of the physiological characteristics after treatment of enzyme replacement therapy (ERT) and oxidative stress. Furthermore, the microfluidic gradient chip developed in this study facilitates the physiological improvement of normal and Fabry disease kidney organoid differentiation and maturation. Therefore, this physiologically enhanced kidney organoid model could be used as a novel in vitro culture system for drug screening applications.
more목차
I. Introduction 6
II. Methods and Materials 9
i. Design and fabrication of microfluidic gradient chip 9
ii. Computational fluid dynamics (CFD) simulation 9
iii. Human induced pluripotent stem cells (hiPSCs) culture and kidney organoids differentiation 10
iv. Generation of Fabry disease kidney organoids 11
v. Preparation on microfluidic chip to culture kidney organoids 14
vi. Immunofluorescence analysis 14
vii. Quantitative RT-PCR 15
viii. Mitochondrial staining and reactive oxygen species (ROS) detection 16
ix. Statistical analysis 17
III. Results and discussions 17
i. Generation of Fabry disease kidney organoids in a microfluidic chip 17
ii. Enhancement of differentiation and maturation of normal kidney organoids in fluidic culture conditions 22
iii. The continuous flow of medium enhances the differentiation and maturation of Fabry disease kidney organoids 26
iv. ERT reduced Gb3 accumulation and attenuated structural and transcriptional changes in Fabry disease kidney organoids 30
v. ERT attenuated oxidative stress and mitochondrial dysfunction in Fabry disease kidney organoids 34
IV. Conclusion 37
V. References 38
