Earticle

초록

영어

Background: Pluripotent stem cells (PSCs) including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) offer the immense therapeutic potential in stem cell-based therapy of degenerative disorders. However, clinical trials of human ESCs cause heavy ethical concerns. With the derivation of iPSCs established by reprogramming from adult somatic cells through the transgenic expression of transcription factors, this problems would be able to overcome. In the present study, we tried to differentiate porcine iPSCs (piPSCs) into endothelial cells (ECs) for stem cell-based therapy of vascular diseases. Methods: piPSCs (OSKMNL) were induced to differentiation into ECs in four differentiation media (APEL-2, APEL-2 + 50 ng/mL of VEGF, EBM-2, EBM-2 + 50 ng/ mL of VEGF) on cultured plates coated with matrigel® (1:40 dilution with DMEM/F-12 medium) for 8 days. Differentiation efficiency of these cells were exanimated using qRT-PCR, Immunocytochemistry, Western blotting and FACS. Results: As results, expressions of pluripotency-associated markers (OCT-3/4, SOX2 and NANOG) were higher observed in all porcine differentiated cells derived from piPSCs (OSKMNL) cultured in four differentiation media than piPSCs as the control, whereas endothelial-associated marker (CD-31) in the differentiated cells was not expressed. Conclusions: It can be seen that piPSCs (OSKMNL) were not suitable to differentiate into ECs in the four differentiation media unlike porcine epiblast stem cells (pEpiSCs). Therefore, it would be required to establish a suitable PSCs for differentiating into ECs for the treatment of cardiovascular diseases.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Induction of porcine induced pluripotent stem cells (piPSCs)
Isolation and culture of swine umbilical vein endothelial cells (SUVECs)
In vitro differentiation of piPSCs into endothelial cells
Alkaline phosphatase (AP) activity
Immunocytochemistry
Quantitative real-time PCR (qRT-PCR)
Western blotting
Flow cytometry (FACS) analysis
Statistical analysis
RESULTS
Morphology of piPSCs cultured for differentiation of endothelial cells
Expressions of pluripotency-associated genes and endothelial-associated gene in differentiated cells derived from piPSCs in differentiation media
Immunocytochemistry of pluripotency-associated markers in differentiated cells derived from piPSCs in differentiation media
Expression of endothelial-associated marker (CD-31) in differentiated cells derived from piPSCs in differentiation media
Flow cytometry of endothelial-associated marker(CD-31) in differentiated cells derived from piPSCs in differentiation media
DISCUSSION
CONCLUSION
REFERENCES

키워드

저자

• In-Won Lee [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University, Jinju 52828, Korea, Division of Applied Life Science, Gyeongsang National University, Jinju 52828, Korea ]
• Hyeon-Geun Lee [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University, Jinju 52828, Korea ]
• Dae-Ky Moon [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University, Jinju 52828, Korea ]
• Yeon-Ji Lee [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University, Jinju 52828, Korea, Division of Applied Life Science, Gyeongsang National University, Jinju 52828, Korea ]
• Bo-Gyeong Seo [ Division of Applied Life Science, Gyeongsang National University, Jinju 52828, Korea, Division of Life Science, College of Natural Sciences, Gyeongsang National University, Jinju 52828, Korea ]
• Sang-Ki Baek [ Gyeongsangnamdo Livestock Experiment Station, Sancheong 52263, 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 ]
• 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

  2019~2025

• 십진분류

  KDC 527 DDC 636