Earticle

초록

영어

Background: Porcine pluripotent stem cells (pPSCs) would provide enormous potential for agriculture and biomedicine. However, authentic pPSCs have not established yet because standards for pPSCs-specific markers and culture conditions are not clear. Therefore, the present study reports comparative pluripotency characteristics in porcine induced pluripotent stem cells (piPSCs) derived from different viral transduction and reprogramming factors [Lenti-iPSCs (OSKM), Lenti-iPSCs (OSKMNL) and Sev-iPSCs (OSKM)]. Methods: Porcine fibroblasts were induced into Lenti-iPSCs (OSKM) and Lenti-iPSCs (OSKMNL) by using Lentiviral vector and Sev-iPSCs (OSKM) by using Sendaiviral vector. Expressions of endogenous or exogenous pluripotency-associated genes, surface marker and in vitro differentiation in between Lenti-piPSCs (OSKM), Lenti-iPSCs (OSKMNL) and Sev-piPSCs (OSKM) were compared. Results: Colonial morphology of Lenti-iPSCs (OSKMNL) closely resembles the naïve mouse embryonic stem cells colony for culture, whereas Sev-iPSCs (OSKM) colony is similar to the primed hESCs. Also, the activity of AP shows a distinct different in piPSCs (AP-positive (+) Lenti-iPSCs (OSKMNL) and Sev-iPSCs (OSKM), but AP-negative (-) LentiiPSCs (OSKM)). mRNAs expression of several marker genes (OCT-3/4, NANOG and SOX2) for pluripotency was increased in Lenti-iPSCs (OSKMNL) and Sev-iPSCs (OSKM), but Sev-iPSCs (OSKM). Interestingly, SSEA-1 of surface markers was expressed only in Sev-iPSCs (OSKM), whereas SSEA-4, Tra-1-60 and Tra-1-81 were positively expressed in Lenti-iPSCs (OSKMNL). Exogenous reprogramming factors continuously expressed in Lenti-iPSCs (OSKMNL) for passage 20, whereas Sev-iPSCs (OSKM) did not express any exogenous transcription factors. Finally, only Lenti-iPSCs (OSKMNL) express the three germ layers and primordial germ cells markers in aggregated EBs. Conclusions: These results indicate that the viral transduction system of reprograming factors into porcine differentiated cells display different pluripotency characteristics in piPSCs.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Culture of porcine fibroblasts
Induction of porcine induced pluripotent stem cells
Determining reprogramming efficiency with alkalinephosphatase
Immunocytochemistry
RT-PCR and quantitative real-time PCR
Cell cycle, population doubling time and karyotypinganalysis
In vitro differentiation
Statistical analysis
RESULTS
Induction efficiency of porcine fibroblasts into porcineinduced pluripotent stem cells by using Lentiviralvector
Induction efficiency of porcine fibroblasts into porcineinduced pluripotent stem cells by using Sendaivirusvector
Comparative expression of endogenous pluripotencyassociatedgenes in between Lenti-piPSCs and SevpiPSCs
Comparative expression of exogenous reprograminggenes in between Lenti-piPSCs and Sev-piPSCs
Comparative expression of pluripotency and surfacemarkers in Lenti-piPSCs and Sev-piPSCs
Cell cycle phase and population doubling time inLenti-piPSCs and Sev-piPSCs
In vitro differentiation of Lenti-piPSCs and SevpiPSCs
DISCUSSION
CONCLUSION
REFERENCES

키워드

저자

• Sang-Ki Baek [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University ]
• In-Won Lee [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University, Division of Applied Life Science, Gyeongsang National University, Jinju 52828, Korea ]
• Yeon-Ji Lee [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University, Division of Applied Life Science, Gyeongsang National University, Jinju 52828, Korea ]
• Bo-Gyeong Seo [ Division of Life Science, College of Natural Sciences, Gyeongsang National University, Division of Applied Life Science, Gyeongsang National University, Jinju 52828, Korea ]
• Jung-Woo Choi [ College of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea ]
• Tae-Suk Kim [ Department of Animal Bioscience, College of Agriculture & Life Sciences, Gyeongsang National University ]
• Cheol Hwangbo [ Division of Applied Life Science, 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

  2019~2026

• 십진분류

  KDC 527 DDC 636