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Protocol for Human Blastoids Modeling Blastocyst Development and Implantation
Published on: August 10, 2022
Development of single blastomeres derived from two-cell embryos produced in vitro in pigs
T Q Dang-Nguyen1, M Kaneda, T Somfai
1National Institute of Livestock and Grassland Science (NILGS), National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki, Japan.
Theriogenology
|March 15, 2011
Summary
Separating single blastomeres from in vitro-produced (IVP) porcine embryos at the two-cell stage significantly increased blastocyst yield. This method offers potential for conserving rare pig breeds by maximizing embryo utilization.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Animal Science
Background:
- In vitro production (IVP) of porcine embryos is crucial for swine reproduction and research.
- Maximizing embryo yield is essential, particularly for conservation efforts of rare breeds.
- Understanding blastomere developmental potential is key to improving IVP efficiency.
Purpose of the Study:
- To investigate the developmental capacity of single blastomeres isolated from two-cell stage porcine embryos produced through IVP.
- To compare blastocyst development rates and cell counts between intact, zona-free, and single blastomere groups.
- To analyze gene expression related to cellular organization and stress response in blastocysts derived from single blastomeres.
Main Methods:
- Porcine embryos were produced in vitro and cultured to the two-cell stage.
- Embryos were divided into three groups: intact (IN), zona-free (ZF), and single blastomere (SB) isolation.
- Blastocyst development rates, cell numbers (inner cell mass and trophectoderm), and expression of specific genes were assessed.
Main Results:
- Blastocyst yield was significantly higher in the SB group (90.2%) compared to IN and ZF groups.
- SB blastocysts had fewer inner cell mass (ICM) and trophectoderm (TE) cells than IN and ZF blastocysts, but ICM:TE ratios were similar.
- Gene expression analysis revealed reduced transcripts for cellular organization (TUBA1, TUBB) in SB blastocysts, but no differences in stress response or glucose metabolism genes.
- Sister blastomere pairs showed a higher tendency to either both develop to blastocysts or both degenerate.
Conclusions:
- Blastomere separation at the two-cell stage enhances blastocyst yield from IVP porcine embryos.
- This technique may be valuable for the conservation of rare pig breeds by increasing the efficiency of embryo transfer.
- Further research is warranted to optimize single blastomere culture conditions and understand long-term developmental implications.
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