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Original Article

Porcine somatic cell nuclear transfer using telomerase reverse transcriptase-transfected mesenchymal stem cells reduces apoptosis induced by replicative senescence

초록

영어

Mesenchymal stem cells (MSCs) have been widely used as donor cells for somatic cell nuclear transfer (SCNT) to increase the efficiency of embryo cloning. Since replicative senescence reduces the efficiency of embryo cloning in MSCs during in vitro expansion, transfection of telomerase reverse transcriptase (TERT) into MSCs has been used to suppress the replicative senescence. Here, TERT-transfected MSCs in comparison with early passage MSCs (eMSCs) and sham-transfected MSCs (sMSCs) were used to evaluate the effects of embryo cloning with SCNT in a porcine model. Cloned embryos from tMSC, eMSC, and sMSC groups were indistinguishable in their fusion rate, cleavage rate, total cell number, and gene expression levels of OCT4, SOX2 and NANOG during the blastocyst stage. The blastocyst formation rates of tMSC and sMSC groups were comparable but significantly lower than that of the eMSC group (p < 0.05). In contrast, tMSC and eMSC groups demonstrated significantly reduced apoptotic incidence (p < 0.05), and decreased BAX but increased BCL2 expression in the blastocyst stage compared to the sMSC group (p < 0.05). Therefore, MSCs transfected with telomerase reverse transcriptase do not affect the overall development of the cloned embryos in porcine SCNT, but enables to maintain embryo quality, similar to apoptotic events in SCNT embryos typically achieved by an early passage MSC. This finding offers a bioengineering strategy in improving the porcine cloned embryo quality.

목차

ABSTRACT
INTRODUCTION
MATERIALS AND METHODS
Chemicals and media
Donor cell preparation
Oocyte preparation
Embryo production
Embryo evaluation of blastocysts stage
Gene expression in blastocyst stage
Statistical analysis
RESULTS
Embryo development: reconstruction and development rate of embryos
Apoptosis and total cell number at the blastocyst stage
Gene expression analysis during the blastocyst stage
DISCUSSION
CONFLICSTS OF INTEREST
AUTHOR CONTRIBUTIONS
AUTHOR’S POSITION AND ORCID NO.
REFERENCES

저자정보

  • Ryounghoon Jeon College of Veterinary Medicine, Gyeongsang National University, Department of Cardiovascular Medicine, Mayo Clinic, Rochester, MN 55905, USA
  • Gyu-Jin Rho College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea

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