Ultrastructure of cloned human blastocyst and first therapeutic cloned Turkish human reconstructed embryos by somatic cell nuclear transfer with intact cytoplasm


Hassa H., Gurer F., Baycu C., Tekin B. , EKER SARIBOYACI A. , Balkose N.

13th World Congress on In-Vitro Fertilization, Assisted Reproduction and Genetics, İstanbul, Türkiye, 26 - 29 Mayıs 2005, ss.91-95 identifier

  • Yayın Türü: Bildiri / Tam Metin Bildiri
  • Cilt numarası:
  • Basıldığı Şehir: İstanbul
  • Basıldığı Ülke: Türkiye
  • Sayfa Sayıları: ss.91-95

Özet

In somatic cell nuclear transfer (SCNT), in an aim to remodelling the chromatin, novel researches are needed to establish the new protocols that include the ultrastructural organizations of developing human reconstructed embryos. For this purpose, in general, one-step enucleation and somatic cell nuclear transfer with cytoplasm to human oocytes has been accomplished. Nuclear and cytoplasmic elements were stained by Papanicolau dye technique as to prevent the parthogenesis. For activation, only electrofusion was applied instead of chemical activation. Moreover, autologous human granulose cells incubated with glucose and 10% human serum albumin (HSA) for three days were used. In control group, same cells were incubated with media having no serum or glucose for three days. Granulose cells of both groups and the cloned human embryos, yielded, from the study group in which cleavage and blastocyst development was present, were examined at electronmicroscopic level. Study group revealed 3 oocytes with remarkable cleavage stage out of 8 reconstructed oocytes. On the other hand, in control group, none of the reconstructed oocytes had a cleavage process. Embryoblasts and trophoblasts of cloned human embryo showed a strong ultrastructural configuration and implantation potential. With regard to granulosa cells, well grown and apoptotic cells were yielded from the study and control groups, respectively. As a result, somatic cell cytoplasm may be of benefit in overcoming the deficiency in centrosome like organelles and/or sperm-related SCNT signal proteins. To accomplish this goal, energy/protein requirements like albumin or glucose should be met. Futhermore, the result of the present study may confer an important contribution to the nucleoli ultrastructural analysis and a successful DNA re-programming, enabling more detailed information regarding ultrastructural details of cloned human embryos.