Investigating effects of primed astrocytes on microglia in Alzheimer’s disease pathology
- Title
- Investigating effects of primed astrocytes on microglia in Alzheimer’s disease pathology
- Author(s)
- Na Yeon Kim
- DGIST Authors
- Na Yeon Kim ; Jinsoo Seo ; Seong-Woon Yu
- Advisor
- 서진수
- Co-Advisor(s)
- Seong-Woon Yu
- Issued Date
- 2024
- Awarded Date
- 2024-02-01
- Type
- Thesis
- Description
- Alzheimer's disease;Priming;Astrocytes;Microglia;Apolipoprotein E;Aβ
- Table Of Contents
-
Ⅰ. Introduction 1
1.1 Neuroinflammatory environment in AD 1
1.2 Innate immune memory : priming effect 2
1.3 Apolipoprotein E4 (ApoE4) and AD 3
1.4 Human-induced pluripotent stem cell model (hiPSC) 3
Ⅱ. Materials and Methods 5
2.1 Cell culture 5
2.1.1 Human-induced pluripotent stem cell (hiPSC) 5
2.1.2 Neural progenitor cells (NPCs) differentiation 5
2.1.3 Astrocyte differentiation 6
2.1.4 Immortalized human microglia 6
2.2 Immune challenge and induction of priming 6
2.3 RNA extraction and cDNA synthesis 6
2.4 quantitative Real-time PCR (qRT-PCR) 7
2.5 Immunocytochemistry 7
2.6 Immunohistochemistry 8
2.7 Cytokine array analysis 8
2.8 Aβ42 uptake assay 9
2.9 Microglial Aβ42 clearance assay 9
2.10 Statistical analysis 10
Ⅲ. Results 11
3.1 Generation of isogenic hiPSC-derived astrocytes and characterization of their identify 11
3.2 Probing priming effect of hiPSC-derived astrocytes by the immune challenge 14
3.3 Aβ42-treated primed astrocytes upregulate the immune response 17
3.4 Aβ42-treated primed astrocytes induce enhancement of Aβ42 clearance of human microglia 20
3.5 APOE4 astrocytes have a priming effect 22
3.6 Primed APOE4 astrocytes enhance Aβ42 uptake but reduce immune response compared with primed APOE3 astrocytes 23
3.7 Aβ42-treated primed APOE4 astrocytes reduce Aβ42 clearance of human microglia 26
3.8 A 9-week-old wild-type (C57BL/6J) male mouse has a priming effect, as in vitro 29
3.9 Confirming the priming effect and Aβ42 clearance of the 7-month-old humanized APOEKI 5XFAD male mouse 33
3.10 Confirming the priming effect and Aβ42 clearance of the 6-month-old humanized APOE KI male mouse 41
Ⅵ. Discussion 44
Ⅴ. References 47
Abstract in Korean 50
List of tables and figures
Table 1. Information of cell lines used in the experiment
Figure 1. Generation of the isogenic hiPSC-derived astrocytes and confirmation of their identify
Figure 2. hiPSC-derived astrocytes have a priming effect
Figure 3. Aβ42-treated primed astrocytes upregulate the immune response
Figure 4. The effect of primed astrocytes on microglia in AD context
Figure 5. APOE4 astrocytes have a priming effect and Aβ42-treated primed APOE4 astrocytes reduce immune response compared with Aβ42-treated primed APOE3 astrocytes
Figure 6. The effect of primed APOE4 astrocytes on microglia in AD context
Figure 7. A wild-type male mouse astrocyte has a priming effect
Figure 8. Confirming the priming effect and Aβ42 clearance of the humanized APOE KI 5XFAD male mouse
Figure 9. Confirming the priming effect and Aβ42 clearance of the humanized APOE KI male mouse
Figure 10. Graphic Summary
- URI
-
http://hdl.handle.net/20.500.11750/48051
http://dgist.dcollection.net/common/orgView/200000724600
- Degree
- Master
- Department
- Department of Brain Sciences
- Publisher
- DGIST
- Related Researcher
-
-
Seo, Jinsoo
- Research Interests iPSC; Alzheimer's disease; Neurodegeneration; Synapse; Neuroscience
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- Appears in Collections:
- Department of Brain Sciences Theses Master
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