Comparison of In Vitro Development of Porcine Embryos Derived from Transfer of Embryonic Germ Cell Nuclei into Oocytes by Electrofusion and Piezo-Driven Microinjection

  • Ahn, Kwang-Sung (Department of Physiology, Dankook University School of Medicine) ;
  • Won, Ji-Young (Department of Physiology, Dankook University School of Medicine) ;
  • Heo, Soon-Young (Department of Physiology, Dankook University School of Medicine) ;
  • Kang, Jee-Hyun (Department of Physiology, Dankook University School of Medicine) ;
  • Shim, Ho-Sup (Department of Physiology, Dankook University School of Medicine)
  • Published : 2007.06.30

Abstract

Embryonic germ (EG) cells are undifferentiated stern cells isolated from cultured primordial germ cells (PGC). These cells share many characteristics with embryonic stem cells including morphology and pluripotency. Undifferentiated porcine EG cell lines demonstrating capacities of differentiation both in vitro and in vivo have been established. Since EG cells can be cultured indefinitely in an undifferentiated state, whereas somatic cells in primary culture are often unstable and have limited lifespan, EG cells may provide inexhaustible source of karyoplasts in nuclear transfer (NT). In this study the efficiencies of NT using porcine EG and fetal fibroblast cells were compared. Two different techniques were used to perform NT. With conventional NT procedure (Roslin method) involving fusion of donor cells with enucleated oocytes, the rates of development to the blastocyst stage in EG and somatic cell NT were 16.8% (59/351) and 14.5% (98/677), respectively. In piezo-driven microinjection (Honolulu method) of donor nuclei into enucleated oocytes, the rates of blastocyst formation in EG and somatic cell NT were 11.9% (15/126) and 9.4% (9/96), respectively. Regardless of NT methods used in this study, EG cell NT gave rise to comparable rate of blastocyst development to somatic cell NT. Overall, EG cells can be used as karyoplast donor in NT procedure, and embryos can be produced by EG cell NT that may be used as an alternative to conventional somatic cell NT.

Keywords

References

  1. Bosch P, Pratt SL, Stice SL (2006): Isolation, characterization, gene modification, and nuclear reprogramming of porcine mesenchymal stem cells. Biol Reprod 74:46-57 https://doi.org/10.1095/biolreprod.105.045138
  2. Colleoni S, Donofrio G, Lagutina I, Duchi R, Galli C, Lazzari G (2005): Establishment, differentiation, electroporation, viral transduction, and nuclear transfer of bovine and porcine mesenchymal stem cells. Cloning Stem Cells 7:154-166 https://doi.org/10.1089/clo.2005.7.154
  3. Faast R, Harrison SJ, Beebe LF, McIlfatrick SM, Ashman RJ, Nottle MB (2006): Use of adult mesenchymal stem cells isolated from bone marrow and blood for somatic cell nuclear transfer in pigs. Cloning Stem Cells 8:166-173 https://doi.org/10.1089/clo.2006.8.166
  4. Hyun SH, Lee GS, Kim DY, Kim HS, Lee SH, Kim S, Lee ES, Lim JM, Kang SK, Lee BC, Hwang WS (2003): Effect of maturation media and oocytes derived from sows or gilts on the development of cloned pig embryos. Theriogenology 59:1641-1649 https://doi.org/10.1016/S0093-691X(02)01211-6
  5. Jin HF, Kumar BM, Kim JG, Song HJ, Jeong YJ, Cho SK, Balasubramanian S, Choe SY, Rho GJ (2007): Enhanced development of porcine embryos cloned from bone marrow mesenchymal stem cells. Int J Dev Biol 51:85-90 https://doi.org/10.1387/ijdb.062165hj
  6. Kawano K, Kato Y, Tsunoda Y (2004): Comparison of in vitro development of porcine nuclear-transferred oocytes receiving fetal somatic cells by injection and fusion methods. Cloning Stem Cells 6: 67-72 https://doi.org/10.1089/1536230041372337
  7. Labosky PA, Barlow DP, Hogan BLM (1994): Mouse embryonic germ (EG) cell lines: transmission through the germline and differences in the methylation imprint of insulin-like growth factor w receptor (Igf2r) gene compared with embryonic stem (ES) cell lines. Development 120:3197-3204
  8. Matsui Y, Toksoz D, Nishikawa S, Williams D, Zsebo K, Hogan BL (1991): Effect of Steel factor and leukaemia inhibitory factor on murine primordial germ cells in culture. Nature 353:750-752 https://doi.org/10.1038/353750a0
  9. Miki H, Inoue K, Kohda T, Honda A, Ogonuki N, Yuzuriha M, Mise N, Matsui Y, Baba T, Abe K, Ishino F, Ogura A (2005): Birth of mice produced by germ cell nuclear transfer. Genesis 41:81-86 https://doi.org/10.1002/gene.20100
  10. Mir B, Tanner N, Chowdhary BP, Piedrahita JA (2003): UP1 extends life of primary porcine fetal fibroblasts in culture. Cloning Stem Cells 5:143-148 https://doi.org/10.1089/153623003322234740
  11. Polejaeva IA, Chen SH, Vaught TD, Page RL, Mullins J, Ball S, Dai Y, Boone J, Walker S, Ayares DL, Colman A, Campbell KHS (2000): Cloned pigs produced by nuclear transfer from adult somatic cells. Nature 407:86-90 https://doi.org/10.1038/35024082
  12. Resnick JL, Bixter LS, Cheng L, Donovan PJ (1992): Long-term proliferation of mouse primordial germ cells in culture. Nature 359:550-551 https://doi.org/10.1038/359550a0
  13. Rideout WM 3rd, Wakayama T, Wutz A, Eggan K, Jackson-Grusby L, Dausman J, Yanagimachi R, Jaenisch R (2000): Generation of mice from wild-type and targeted ES cells by nuclear cloning. Nat Genet 24:109-110 https://doi.org/10.1038/72753
  14. Roh S, Hwang WS (2002): In vitro development of porcine parthenogenetic and cloned embryos: comparison of oocyte-activating techniques, various culture systems and nuclear transfer methods. Reprod Fertil Dev 14:93-99 https://doi.org/10.1071/RD01090
  15. Shim H, Gutierrez-Adan A, Chen LR, BonDurant RH, Behboodi E, Anderson GB (1997): Isolation of pluripotent stem cells from cultured porcine primordial germ cells. Biol Reprod 57:1089-1095 https://doi.org/10.1095/biolreprod57.5.1089
  16. Wakayama T, Perry AC, Zuccotti M, Johnson KR, 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
  17. Wakayama T, Rodriguez I, Perry AC, Yanagimachi R, Mombaerts P (1999): Mice cloned from embryonic stem cells. Proc Natl Acad Sci USA 96:14984-14989
  18. Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KHS (1997): Viable offspring derived from fetal and adult mammalian cells. Nature 385:810-813 https://doi.org/10.1038/385810a0
  19. Yamazaki Y, Mann MR, Lee SS, Marh J, McCarrey JR, Yanagimachi R, Bartolomei MS (2003): Reprogramming of primordial germ cells begins before migration into the genital ridge, making these cells inadequate donors for reproductive cloning. Proc Natl Acad Sci USA 100:12207-12212
  20. Zhu H, Craig JA, Dyce PW, Sunnen N, Li J (2004): Embryos derived from porcine skin-derived stem cells exhibit enhanced preimplantation development. Biol Reprod 71:1890-1897 https://doi.org/10.1095/biolreprod.104.032227