Earticle

초록

영어

Background: Endothelial cells (EC) that make up the inner wall of blood vessels play an important role in angiogenesis and vascular recovery. Cardiovascular disease caused by dysfunction of ECs has been reported as a major cause of death worldwide. Despite significant research so far, the underlying mechanism of dysfunction of ECs in cardiovascular disease progression is not yet fully understood. Although therapeutic transplantation of autologous ECs is limited due to lack of cell availability, adiposederived stem cells (ADSCs), known for their ease of procurement and high potential for differentiation, will provide promising solutions to generate autologous ECs. Methods: This study investigated the optimal differentiation of ADSCs into ECs under EBM-2 culture conditions supplemented with VEGF and BMP-4 in hypoxia (2% O2). Results: During 14 days of in vitro differentiation, cells cultured in EBM-2 supplemented with VEGF showed the characteristics of early vascular ECs and some cells adopted polygonal forms. Conversely, cells cultured in EBM-2 and hypoxia supplemented with both VEGF and BMP-4 differentiated into the typical cobblestone morphology that appears in vascular ECs. As a result of immunostaining against the vascular ECs marker CD-31, CD-31 expression was increased under EBM-2 culture conditions with VEGF and VEGF + BMP-4 in hypoxia, but expression was insufficient in normal oxygenation (21% O2). In the flow cytometry analysis, high expression of CD-31 expression was observed under conditions including both VEGF and BMP-4 of hypoxia. Interestingly, in gene expression, the pluripotency marker OCT-3/4 was significantly reduced under hypoxic conditions, but SOX2 and NANOG expression were higher than under normal oxygen conditions. However, CD-31 expression was significantly higher under differentiation conditions in which VEGF and BMP-4 were added under hypoxia conditions. In a functional analysis, CD-31-positive ADSC-derived ECs differentiated under hypoxia had excellent tube formation and Dil-Ac-LDL uptake, which are important for vascular repair and function. Conclusions: These findings confirmed the therapeutic usefulness of ECs derived from ADSC for the treatment of cardiovascular disease due to the synergy effect of hypoxia and BMP-4.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Isolation and culture of swine umbilical vein endothelial cells (SUVEC)
Isolatiuon and culture of porcine adipose-derived stem cells (pADSCs)
In vitro differentiation to endothelial cells (ECs)
Immunocytochemistry
Quantitative real-time PCR (qPCR)
Flow cytometry (FACS) analysis
Magnetic activated cell sorting (MACS)
Tube formation assay
Dil-ac-LDL uptake assay
Statistical analysis
RESULTS
Morphological changes during in vitro differentiation from hypoxic conditions to ECs
Expression of CD-31 after in vitro differentiation of pADSCs under hypoxic conditions
Expression of pluripotency-related genes and endothelial gene of pADSCs differentiated in hypoxia
Flow cytometry validation of CD-31 expression under hypoxic culture conditions
Formation of capillary-like structures of pADSC-derivedendothelial cells
Assessment of function via Dil-Ac-LDL uptake in pADSCs-derived endothelial cells
DISCUSSION
CONCLUSION
REFERENCES

키워드

저자

• In-Won Lee [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University/Division of Applied Life Science (BK21 Four), Gyeongsang National University, Jinju 52828, Korea ]
• Dong-Ju Shin [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University/Division of Applied Life Science (BK21 Four), Gyeongsang National University, Jinju 52828, Korea ]
• Tae-Suk Kim [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University, Jinju 52828, Korea ]
• Cheol Hwangbo [ Division of Life Science, College of Natural Sciences, Gyeongsang National University, Jinju 52828, Korea ]
• Jung-Woo Cho [ College of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea ]
• Sang-Hee Lee [ College of Animal Life Science, Kangwon National University/School of ICT, University of Tasmania, Hobart 7005, Australia ] Corresponding Author
• Joon-Hee Lee [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University/Institute of Agriculture & Life Science, College of Agriculture & Life Sciences, Gyeongsang National University, Jinju 52828, Korea ] Corresponding Author

참고문헌

발행기관

간행물

관심저널 등록

• 간행물명

  Journal of Animal Reproduction and Biotechnology

• 간기

  계간

• pISSN

  2671-4639

• eISSN

  2671-4663

• 수록기간

  2019~2026

• 십진분류

  KDC 527 DDC 636