Bidirectional transcriptome analysis of rat bone marrow-derived mesenchymal stem cells and activated microglia in an in vitro coculture system
- 주제(키워드) bone marrow-derived mesenchymal stem cells , inflammation , microglia , migration , transcriptome analysis
- 발행기관 아주대학교
- 지도교수 이광
- 발행년도 2020
- 학위수여년월 2020. 8
- 학위명 박사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000030178
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Microglia contribute to the pathogenesis of brain diseases by regulating of neuroinflammation. Thus, targeting of neuroinflammation triggered by activated microglia in brain diseases has become a promising curative strategy. Bone marrow-derived mesenchymal stem cells (BM-MSCs) have been shown to have therapeutic effects, resulting from the regulation of inflammatory conditions in the brain. In this study, Gene expression pattern in rat BM-MSCs (rBM-MSCs) cocultured with lipopolysaccharide- (LPS-) stimulated primary rat microglia were investigated using microarray analysis and the functional relationships were evaluated through Ingenuity Pathway Analysis (IPA). The effects of rBM-MSC on LPS-stimulated microglia were also assessed using a reverse coculture system and the same transcriptomic analysis. In rBM-MSCs, 67 genes were differentially expressed, which were highly related with migration of cells, compared to control. The gene network was predicted using IPA and LPS-stimulated primary rat microglia increase the migration of rBM-MSCs was validated by experiments. Reversely, expression patterns of the transcriptome in LPS-stimulated primary rat microglia cocultured with rBM-MSCs were changed. Results showed that 64 genes were altered, which were highly related with inflammatory response, compared to absence of rBM-MSCs. In the same procedure with the aforementioned, the prediction of the gene network and experimental validation showed that rBM-MSCs decrease the inflammatory response of LPS-stimulated primary rat microglia. These indicate that LPS-stimulated microglia increase the migration of rBM-MSCs and that rBM-MSCs reduce the inflammatory activity in LPS-stimulated microglia. The results of this study show complex mechanisms underlying the interaction between rBM-MSCs and activated microglia and may be supportive for the advance of stem cell therapy for brain diseases. This study is based on a previously published report
more목차
1. Introduction 1
2. Materials and methods 5
2.1. Isolation and maintenance of rat bone marrow-derived mesenchymal stem cells (rBM-MSCs) 5
2.2. Rat microglia primary cultures 7
2.3. Coculture of LPS-stimulated or non-stimulated microglia and rBM-MSCs 9
2.4. Total RNA isolation and microarray analysis 9
2.5. Transcriptomic analysis using Ingenuity Pathway Analysis 10
2.6. Quantitative real-time PCR (qPCR) 14
2.7. Migration assay 17
2.8. Statistical analysis 17
3. Results 18
3.1. Cellular movement-related transcriptomic changes in rBM-MSCs cocultured with LPS-stimulated microglia 18
3.2. Functional prediction of transcriptomic networks in rBM-MSCs cocultured with LPS-stimulated microglia 23
3.3. Increased migratory activity in rBM-MSCs cocultured with LPS-stimulated microglia 28
3.4. Transcriptomic analysis in LPS-stimulated microglia cocultured with rBM-MSCs 30
3.5. Functional prediction of transcriptomic networks and reduced inflammatory response in LPS-stimulated microglia cocultured with rBM-MSCs 34
4. Discussion 41
CONCLUSION 44
REFERENCES 45
