Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
권호 표지

In Vitro Development of Interspecies Nuclear Transfer Embryos: Effects of Culture Systems

  • Roh Sangho (Section of Oral and Craniofacial Biotechnology Research, Dental Research Institute and College of Dentistry, Seoul National University)
  • Published : 2004.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

Porcine fibroblasts were transferred into enucleated bovine oocytes for the interspecies nuclear transfer (NT). After NT, the embryos were cultured in three different culture systems. The media used for the experiment were CR1aa and NCSU23. The culture systems used for the experiment were: 1. Culture in CR1aa for 7 days (CR). 2. Culture in CR1aa for 2 days and subsequently in NCSU23 for 5 days (CR-NC). 3. Culture in NCSU23 for 7 days (NC). Bovine (intraspecies) NT group was used as a control. The oocytes in bovine NT group were treated the same as interspecies NT embryos except using bovine fibroblasts as nuclear donors. Regardless of their nuclear origin (interspecies vs bovine), the embryos in CR (68.4% vs 77.2%) and CR-NC (67.8% vs 70.5%) showed better developmental competence to the 2-cell stage (p<0.05) than those in NC (41.0% vs 10.0%). Bovine NT embryos in CR-NC did not develop over the 4-cell stage after the medium replacement, while interspecies NT embryos in CR-NC continued to develop and could reach over the 8-cell stage (12.2%). Blastocysts were only found in bovine NT group (17.4%), but no blastocyst was found in interspecies NT group. This study suggests that the development of interspecies NT embryos mostly depends on their recipient cytoplasm during the culture in vitro.

Keywords

References

  1. Anderson JE, Matteri RL, Abeydeera LR, Day BN. and Prather RS (2001): Degradation of maternal cdc25c during the meternal to zygotic transition is dependent upon embryonic transcription. Mol Reprod Dev 60:181-188
  2. Bachvarova R, Cohen EM, De Leon V, Tokunaga K, Sakiyama S, Paynton BV (1989): Amounts and modulation of actin mRNAs in mouse oocytes and embryos. Development 106:561-565
  3. Barnes FL, First NL (1991): Embryonic transcription in in vitro cultured bovine embryos. Mol Reprod Dev 29:117-123 https://doi.org/10.1002/mrd.1080290205
  4. Cummins, J (1998): Mitochondrial DNA in mammalian reproduction. Rev Reprod 3:172-182 https://doi.org/10.1530/ror.0.0030172
  5. Dominko T, Mitalipova M, Haley B, Beyhan Z, Memili E, McKusick B, First NL (1999): Bovine oocyte cytoplasm supports development of embryos produced by nuclear transfer of somatic cell nuclei from various mammalian species. Biol Reprod 60: 1496-1502 https://doi.org/10.1095/biolreprod60.6.1496
  6. Eyestone WH, First, NL (1986): A study of the 8- to 16-cell developmental block in bovine embryos cultured in vitro. Theriogenology 25:152 https://doi.org/10.1016/0093-691X(86)90206-2
  7. Henrion G, Adeline B, Renard JP, Duranthon V (1997): Identification of maternal transcripts that progressively disappear during the cleavage period of rabbit embryos. Mol Reprod Dev 47:353-362 https://doi.org/10.1002/(SICI)1098-2795(199708)47:4<353::AID-MRD1>3.0.CO;2-J
  8. Kitiyanant Y, Saikhun J, Chaisalee B, White KL, Pavasuthipaist K (2001): Somatic cell cloning in Buffalo (Bubalus bubalis): Effects of interspecies cytoplasmic recipients and activation procedures. Cloning Stem Cells 3:97-104
  9. Lanza RP, Cibelli JB, Diaz F, Morales CT, Farin PW, Farin CE, Hammer CJ, West MD, Damiani P (2000): Cloning of an endangered species (Bas gaurus) using interspecies nuclear transfer. Cloning 2:79-90
  10. Lawitts JA, Biggers JD (1991): Optimization of mouse embryo culture media using simplex methods. J Reprod Fertil 91:543-556 https://doi.org/10.1530/jrf.0.0910543
  11. Miyoshi K, Kono T, Niwa K (1997): Stage-dependent development of rat I-cell embryos in a chemically defined medium after fertilization in vivo and in vitro. Biol Reprod 56:180-185 https://doi.org/10.1095/biolreprod56.1.180
  12. Petters RM, Wells KD (1993): Culture of pig embryos. J Reprod Fertil Suppl 48: 61-73
  13. Prather RS (1993): Nuclear control of preimplantation development in the domestic pig. 4th International conference on pig reproduction. J Reprod Fertil Suppl 48:17-29
  14. Rosenkrans CF Jr, First NL (1991): Culture of bovine zygote to blastocyst stage: effects of amino acids and vitamins. Theriogenology 35:266 https://doi.org/10.1016/0093-691X(91)90242-6