Earticle

초록

영어

The faulty regulation of imprinting gene lead to the abnormal development of reconstructed embryo after nuclear transfer. However, the correlation between the imprinting status of donor cell and preimplantation stage of embryo development is not yet clear. In this study, to determine this correlation, we used the porcine spermatogonial stem cell (pSSC) and fetal fibroblast (pFF) as donor cells. As the results, the isolated cells with laminin matrix selection strongly expressed the GFRα-1 and PLZF genes of SSCs specific markers. The pSSCs were maintained to 12 passages and positive for the pluripotent marker including OCT4, SSEA1 and NANOG. The methylation analysis of H19 DMR of pSSCs revealed that the zinc finger protein binding sites CTCF3 of H19 DMRs displayed an androgenic imprinting pattern (92.7%). Also, to investigate the reprogramming potential of pSSCs as donor cell, we compared the development rate and methylation status of H19 gene between the reconstructed embryos from pFF and pSSC. This result showed no significant differences of the development rate between the pFFs (11.2±0.8%) and SSCs (13.3±1.1%). However, interestingly, while the CTCF3 methylation status of pFF-NT blastocyst was decreased (36.3%), and the CTCF3 methylation status of pSSC-NT blastocyst was maintained. Therefore, this result suggested that the genomic imprinting status of pSSCs is more effective than that of normal somatic cells for the normal development because the maintenance of imprinting pattern is very important in early embryo stage.

목차

ABSTRACT
 INTRODUCTION
 MATERIALS AND METHODS
  Isolation and Culture of Porcine SSCs
  Immunohistochemistry
  Immunocytochemistry
  Oocyte Retrieval and In Vitro Maturation
  Somatic Cell Nuclear Transfer
  Bisulfate Treatment
  PCR Amplification and Sequencing of Bisulfite-treatedGenomic DNA
  Reverse Transcriptase-polymerase Chain Reaction
  Statistical Analyses
 RESULTS
  The Histological Analysis of Porcine Testis
  The Isolation and Characterization of pSSCs
  Analysis of the Methylation Status in pSSCs and pSSCNTEmbryo
 DISCUSSION
 REFERENCES

키워드

저자

• Hyun Seung Lee [ Department of Animal Biotechnology, Animal Resources Research Center/Bio-Organ Research Center, Konkuk University, Seoul 143-701, Korea ]
• Sung Ho Lee [ Department of Animal Biotechnology, Animal Resources Research Center/Bio-Organ Research Center, Konkuk University, Seoul 143-701, Korea ]
• Mukesh Kumar Gupta [ Department of Animal Biotechnology, Animal Resources Research Center/Bio-Organ Research Center, Konkuk University, Seoul 143-701, Korea ]
• Sang Jun Uhm [ Department of Animal Biotechnology, Animal Resources Research Center/Bio-Organ Research Center, Konkuk University, Seoul 143-701, Korea ]
• Hoon Taek Lee [ Department of Animal Biotechnology, Animal Resources Research Center/Bio-Organ Research Center, Konkuk University, Seoul 143-701, Korea ] Corresponding author

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간행물

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• 간행물명

  Reproductive & developmental biology

• 간기

  계간

• pISSN

  1738-2432

• eISSN

  2288-0151

• 수록기간

  1977~2018

• 십진분류

  KDC 527 DDC 636