Flexible nanodotsensor for drug evaluation of human glioblastoma-midbrain assembloid
- 주제어 (키워드) Glioblastoma , Midbrain , Adhesive hydrogel , flexible nanodot sensor , Electrophysiological signals , Drug evaluation , Neural connectivity.
- 발행기관 서강대학교 일반대학원
- 지도교수 최정우
- 발행년도 2024
- 학위수여년월 2024. 8
- 학위명 석사
- 학과 및 전공 일반대학원 화공생명공학과
- 실제 URI http://www.dcollection.net/handler/sogang/000000079207
- UCI I804:11029-000000079207
- 본문언어 영어
- 저작권 서강대학교 논문은 저작권 보호를 받습니다.
초록 (요약문)
Glioblastoma multiforme (GBM), a highly malignant brain tumor, significantly impacts the central nervous system and is associated with low survival rates. Despite various organoid models mimicking human GBM, weak signals from existing brain organoids challenge drug screening. To address this gap, we developed a brain tumor- midbrain assembloid using an adhesive hydrogel incorporating neurotrophic factor-3 (NT3) and gold nanoparticles (GNP), and improved drug response evaluation by developing a sensitive flexible nanodot sensor. The NT3/GNP-incorporated adhesive hydrogel showed a 1.78-fold increase in electrical conductivity and a 1.93-fold decrease in resistance compared to the control hydrogel. Electrophysiological signals analyzed using multi- electrode array (MEA) platforms demonstrated a 2-fold increase in spike activity in the NT3/GNP-incorporated hydrogel assembloid, indicating enhanced neural connectivity. For drug evaluation, electrophysiological recordings were performed using the flexible nanodot sensor. The GBM-midbrain assembloid treated with Everolimus (0.1 μM) for one week showed a significant decrease in electrophysiological signals, demonstrating the sensor's effectiveness in assessing drug responses. These findings highlight the potential of the NT3/GNP-incorporated adhesive hydrogel in investigating GBM organoid interactions and the utility of the flexible nanodot sensor in drug evaluation.
more목차
1. Introduction 01
2. Experimental details 06
2.1. M a t e r i a l s 06
2.2. hiPSC cu l ture 09
2.3. Generation of midbrain and cerebral organoids 09
2.4. Transfection and generation of GBM organoid 12
2.5. Synthesis of the NT3/GNP-incorporated adhesive hydrogel 13
2.6. Formation of glioblastoma-midbrain assembloid 15
2.7. Tissue clearing and immunofluorescence staining 16
2.8. RNA isolation and cDNA preparation 17
2.9. Analysis of quantitative RT-PCR 17
2.10. Electrophysiological activity analysis 19
2.11. Construction of flexible nanodot electrode 20
2.12. Fabrication of nanodot-patterned concave sensor 21
2.13. Electrochemical characterization 21
2.14. Drug evaluation of assembloid using nanodot elect rode 22
3. Results and Discussion 23
3.1. Generation of midbrain organoids from hiPSC 23
3.2. Generation of cerebral and glioblastoma organoids 26
3.3. Synthesis and properties of the NT3/GNP-incorporated hydrogel 30
3.4. Formation of the GBM-MB assembloid 34
3.5. Fabrication of f lexible nanodot sensor 37
3.6. Electrophysiological recordings of assembloid 39
4. Conclusion 42
5. Reference 43
