DOI QR코드

DOI QR Code

Comparison of Developmental Efficiency of Murine Somatic Cell Nuclear Transfer Protocol

  • Moon, Jeonghyeon (Cellular Reprogramming and Embryo Biotechnology Laboratory, Dental Research Institute, BK21, Seoul National University School of Dentistry) ;
  • Jung, Miran (ET Biotech Co. Ltd.) ;
  • Roh, Sangho (Cellular Reprogramming and Embryo Biotechnology Laboratory, Dental Research Institute, BK21, Seoul National University School of Dentistry)
  • Received : 2017.08.04
  • Accepted : 2017.09.17
  • Published : 2017.09.30

Abstract

The Somatic cell nuclear transfer (SCNT) method can be applied to various fields such as species conservation, regenerative medicine, farming industries and drug production. However, the efficiency using SCNT is very low for many reasons. One of the troubles of SCNT is that it is highly dependent on the researcher's competence. For that reason, four somatic cell nuclear injection methods were compared to evaluate the effect of hole-sealing process and existence of cytochalasin B (CB) on efficiency of murine SCNT protocol. As a results, the microinjection with the hole-sealing process, the oocyte plasma membrane is inhaled with injection pipette, in HCZB with CB was presented to be the most efficient for the reconstructed in SCNT process. In addition, we demonstrated that the oocytes manipulated in Hepes-CZB medium (HCZB) with CB does not affect the developmental rate and the morphology of the blastocyst during the pre-implantation stage. For this reason, we suggest the microinjection involving hole-sealing in HCZB with CB could improve SCNT process efficiency.

Keywords

References

  1. Munsie MJ, Michalska AE, O'Brien CM, Trounson AO, Pera MF, and Mountford PS. 2000. Isolation of pluripotent embryonic stem cells from reprogrammed adult mouse somatic cell nuclei. See comment in PubMed Commons belowCurr Biol. 10:989-992.
  2. Byrne JA, Pedersen DA, Clepper LL, Nelson M, Sanger WG, Gokhale S, Wolf DP, and Mitalipov SM. 2007. Producing primate embryonic stem cells by somatic cell nuclear transfer. Nature 450:497-502. https://doi.org/10.1038/nature06357
  3. Chung YG, Eum JH, Lee JE, Shim SH, Sepilian V, Hong SW, Lee Y, Treff NR, Choi YH, Kimbrel EA, Dittman RE, Lanza R, and Lee DR. 2014. Human somatic cell nuclear transfer using adult cells. Cell Stem Cell 14:777-780. https://doi.org/10.1016/j.stem.2014.03.015
  4. Yamada M, Johannesson B, Sagi I, Burnett LC, Kort DH, Prosser RW, Paull D, Nestor MW, Freeby M, Greenberg E, Goland RS, Leibel RL, Solomon SL, Benvenisty N, Sauer MV, and Egli D. 2014. Human oocytes reprogram adult somatic nuclei of a type 1 diabetic to diploid pluripotent stem cells. Nature 510:533-536. https://doi.org/10.1038/nature13287
  5. Campbell KH, McWhir J, Ritchie WA, and Wilmut I. 1996. Sheep cloned by nuclear transfer from a cultured cell line. Nature 380:64-66. https://doi.org/10.1038/380064a0
  6. Wakayama T, Perry AC, Zuccotti M, Johnson KR, and Yanagimachi R. 1998. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 394:369-374. https://doi.org/10.1038/28615
  7. Kato Y, Tani T, Sotomaru Y, Kurokawa K, Kato J, Doguchi H, Yasue H, and Tsunoda Y. 1998. Eight calves cloned from somatic cells of a single adult. Science 282:2095-8. https://doi.org/10.1126/science.282.5396.2095
  8. Kishigami S, Wakayama S, Thuan NV, Ohta H, Mizutani E, Hikichi T, Bui HT, Balbach S, Ogura A, Boiani M, and Wakayama T. 2006. Production of cloned mice by somatic cell nuclear transfer. Nat. Protoc. 1:125-138. https://doi.org/10.1038/nprot.2006.21
  9. Markoulaki S, Meissner A, and Jaenisch R. 2008. Somatic cell nuclear transfer and derivation of embryonic stem cells in the mouse. Methods 45:101-114. https://doi.org/10.1016/j.ymeth.2008.04.002
  10. Kishigami S, Mizutani E, Ohta H, Hikichi T, Thuan NV, Wakayama S, Bui HT, and Wakayama T. 2006. Significant improvement of mouse cloning technique by treatment with trichostatin A after somatic nuclear transfer. Biochem. Biophys. Res. Commun. 340:183-189. https://doi.org/10.1016/j.bbrc.2005.11.164
  11. Terashita Y, Wakayama S, Yamagata K, Li C, Sato E, and Wakayama T. 2012. Latrunculin A can improve the birth rate of cloned mice and simplify the nuclear transfer protocol by gently inhibiting actin polymerization. Biol. Reprod. 86:180.
  12. Matoba S, Liu Y, Lu F, Iwabuchi KA, Shen L, Inoue A, and Zhang Y. 2014. Embryonic development following somatic cell nuclear transfer impeded by persisting histone methylation. Cell 159:884-895. https://doi.org/10.1016/j.cell.2014.09.055
  13. A.J. French, C.A. Adams, L.S. Anderson, J.R. Kitchen, M.R. Hughes, and S.H. Wood. 2008. Development of human cloned blastocysts following somatic cell nuclear transfer with adult fibroblasts. Stem Cells 26:485-493. https://doi.org/10.1634/stemcells.2007-0252
  14. Noggle S, Fung HL, Gore A, Martinez H, Satriani KC, Prosser R, Oum K, Paull D, Druckenmiller S, Freeby M, Greenberg E, Zhang K, Goland R, Sauer MV, Leibel RL, and Egli D. 2011. Human oocytes reprogram somatic cells to a pluripotent state. Nature 478:70-75. https://doi.org/10.1038/nature10397
  15. Egli D, and Eggan K. 2006. Nuclear transfer into mouse oocytes. J. Vis. Exp. 116.
  16. MacLean-Fletcher S, and Pollard TD. 1980. Mechanism of action of cytochalasin B on actin. Cell 20:329-341. https://doi.org/10.1016/0092-8674(80)90619-4