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Convenient Monitoring System of Intracellular microRNA Expression during Adipogenesis via Mechanical Stimulus-Induced Exocytosis of Lipovesicular miRNA Beacon

 Seungmin Han  ;  Byunghoon Kang  ;  Eunji Jang  ;  Jisun Ki  ;  Eunjung Kim  ;  Mun-Young Jeong  ;  Yong-Min Huh  ;  Hye-Young Son  ;  Seungjoo Haam 
 Advanced Healthcare Materials, Vol.7(5) : 1701019, 2018 
Journal Title
 Advanced Healthcare Materials 
Issue Date
differentiation ; exocytosis ; liposome ; miRNA sensing ; real-time monitoring
Noninvasive investigation of microRNAs (miRNAs) expression, which is deeply related to biological phenomena such as stem cell differentiation, in culture soup is particularly useful for monitoring of stem cell differentiation without phototoxicity of living cells, especially when cell morphologies remain unchanged during differentiation. However, real-time detection of miRNA in culture soup is not recommended because of insufficient miRNA amounts in culture soup. In this study, a convenient method is introduced for real-time assessing intracellular miRNA in culture soup by using lipovesicular miRNA beacon (Lipo-mB) and mechanical stimulus-mediated exocytosis. Pipetting-harvest of culture soup induces exocytosis-secretion of fluorescence signal of Lipo-mB from cytoplasm into culture soup. To demonstrate this method, Lipo-mB is applied for monitoring of adipogenesis by analyzing the expression levels of various intracellular miRNAs, which are related to adipogenesis regulators. The fluorescence intensity profile of the culture soup is correlated with the quantitative reverse-transcription-polymerase chain reaction data and absorbance of Oil Red O staining. These results demonstrate that Lipo-mB can successfully monitor stem cell differentiation by sensing changes in miRNA expression from culture soup of living cells. Lipo-mB can be further developed as an accurate sensing system for analyzing subtle differences in genotype, even when changes in phenotype cannot be observed.
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1. College of Medicine (의과대학) > BioMedical Science Institute (의생명과학부) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Radiology (영상의학교실) > 1. Journal Papers
Yonsei Authors
손혜영(Son, Hye Yeong) ORCID logo https://orcid.org/0000-0001-5977-6784
허용민(Huh, Yong Min) ORCID logo https://orcid.org/0000-0002-9831-4475
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