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Embryo Aggregation in Pig Improves Cloning Efficiency and Embryo Quality.

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Embryo aggregation in pigs improves cloned embryo development and quality. Aggregating three cloned embryos enhances blastocyst formation and reduces DNA fragmentation, indicating better cell reprogramming and lower apoptosis.

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Area of Science:

  • Reproductive Biology
  • Developmental Biology
  • Animal Science

Background:

  • Cloned embryo development in pigs often faces challenges with quality and developmental rates.
  • Improving in vitro development and embryo quality is crucial for efficient production of cloned livestock.

Purpose of the Study:

  • To investigate the effects of cloned embryo aggregation on in vitro development and quality in porcine embryos.
  • To assess the impact of aggregation on blastocyst formation, cell number, DNA fragmentation, and gene expression related to pluripotency, apoptosis, and methylation.

Main Methods:

  • Zona-free reconstructed cloned porcine embryos were cultured using the 'well of the well' system.
  • Embryos were grouped as single (1x non-aggregated) or aggregated (3x per microwell).
  • Evaluated blastocyst diameter, cell number, DNA fragmentation, and expression of key genes (Oct4, Nanog, Klf4, Igf2, Bax, Dnmt1, Cdx2, Bcl-xl).

Main Results:

  • Aggregation of three embryos significantly increased blastocyst formation rate and blastocyst diameter.
  • DNA fragmentation levels were significantly decreased in aggregated cloned blastocysts compared to non-aggregated ones.
  • Expression of pluripotency (Oct4, Klf4), growth (Igf2), apoptosis (Bax), and methylation (Dnmt1) genes increased, while Nanog expression was unaffected. Cdx2 and Bcl-xl were undetectable.

Conclusions:

  • Embryo aggregation in pigs positively influences cloned embryo development and quality.
  • Aggregation may enhance cloned embryo viability by reducing apoptosis and improving cellular reprogramming.
  • This technique shows potential for improving the efficiency of porcine cloning.