Mass Production of Exosomes from Human Stem Cells by Inducing Cellular Apoptosis
- 주제(키워드) Exosomes , Adipose tissue-derived mesenchymal stem (LMSCs) , mesenchymal stem cells (MSCs) , pre-conditioning , characterization , large-scale isolation.
- 발행기관 아주대학교
- 지도교수 윤승현
- 발행년도 2022
- 학위수여년월 2022. 2
- 학위명 박사
- 학과 및 전공 일반대학원 의학과
- 실제URI http://www.dcollection.net/handler/ajou/000000031487
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Abstract Exosomes are nano-sized extracellular vesicles that have been extensively investigated for their role in cell-to-cell communication and disease progression. Owing to its inherent properties of low size, high safety profile, and excellent cellular penetrability, they are favorable candidates for many therapeutic purposes including targeted therapy, drug delivery and vaccine platform. While there is exponential rise in research exploring potential therapeutic effect of exosomes, they are still in the preliminary stage of development towards a clinical therapy. Exosome being a cell secretory product, to exploit them as scalable, cGMP-compliant manufacturing level is a major challenge at present. Here I present a strategy for the mass isolation of exosome using conditioned media of adipose derived mesenchymal stem cells. This approach is ultracentrifugation based enhanced secretion of apoptotic cell derived exosome where isolated exosomes were characterized by NTA, electron microscopy and Exoview. The purpose of this paper is to overcome the existing barrier of low exosome yield thus, enabling large-scale exosome manufacturing which may lead to rapid translation of exosome-based therapy into clinical settings.
more목차
I. INTRODUCTION 1
II. MATERIAL AND METHODS 4
A. MSCs isolation and culture 4
B. Amplification of mesenchymal stem cells 4
C. Culturing of mesenchymal stem cells in conditioned media 5
D. Isolation of exosomes from conditioned mesenchymal stem cell cultivating medium 5
E. Detection and quantification of exosomes in MSC-conditioned medium 6
F. Effect of DNase I treatment 9
III. RESULTS 10
A. Adipose derived stem cell (LMSC) characterization 10
B. Characterization of Isolated Exosomes from MSC-conditioned medium 10
C. DNase I treatment enhances purity of exosomes 17
D. Protein profile 18
E. Data validation 22
IV. DISCUSSION 24
V. CONCLUSION 33
REFERENCES 34
