한국수정란이식학회지 (Journal of Embryo Transfer)

  • 제26권2호
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  • Pages.111-115
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  • 2011
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  • 2508-755X(pISSN)
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  • 2288-0178(eISSN)
한국수정란이식학회 (The Korean Society of Embryo Transfer)
권호 표지

Comparison of the Methods of Zona Pellucida Removal and Inner Cell Mass Isolation for the Generation of Parthenogenetic Embryonic Stem Cells in HanWoo Cattle

  • Kim, Dae-Hwan (Cellular Reprogramming and Embryo Biotechnology Laboratory, Dental Research Institute and CLS21, Seoul National University School of Dentistry) ;
  • Park, Sang-Kyu (Cellular Reprogramming and Embryo Biotechnology Laboratory, Dental Research Institute and CLS21, Seoul National University School of Dentistry) ;
  • Kim, Se-Woong (Cellular Reprogramming and Embryo Biotechnology Laboratory, Dental Research Institute and CLS21, Seoul National University School of Dentistry) ;
  • Jung, Yeon-Gil (ET Biotech) ;
  • Roh, Sang-Ho (Cellular Reprogramming and Embryo Biotechnology Laboratory, Dental Research Institute and CLS21, Seoul National University School of Dentistry)
  • 투고 : 2011.04.15
  • 심사 : 2011.04.27
  • 발행 : 2011.06.30
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초록

In general, zona pellucida (ZP) of the blastocyst has to be removed first, then either isolated the inner cell mass (ICM) or ZP-removed whole blastocyst, which is then cultured on the feeder layer to induce ICM outgrowth for the generation of embryonic stem cells (ESC). However, it is unclear whether ICM isolation before seeding on feeder layer is beneficial or not because the interaction between ICM and trophoblasts may affect cellular growth and/or pluripotency during the culture on the feeder. In the present study, two ZP removal methods (mechanically by splitting with a 28-gauge needle versus chemically by the treatment of acid-Tyrode's solution) and two ICM isolation methods (ZP-free whole blastocyst seeding versus mechanical isolation of ICM) were evaluated for the efficient isolation and culture of putative parthenogenetic bovine ESC. The number of maintained outgrown colonies was counted in each experimental group. As the result, mechanical removal of ZP with a needle and followed by whole ZP-free blastocyst seeding on feeder cells tended to attach more on the feeder layer and resulted in more outgrown colonies with its simple and less time-costing benefits. Currently we are generating ESC lines in HanWoo cattle by using this method for initial outgrowth of the parthenogenetic bovine blastocysts.

키워드

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