Cellular reprogramming by single-cell fusion with mouse embryonic stem cells in pig

Area of Science:

• Cell Biology
• Stem Cell Research
• Reproductive Biology

Background:

• Cellular reprogramming via somatic cell fusion is crucial for regenerative medicine but suffers from low efficiency.
• Traditional methods like polyethylene glycol (PEG) fusion have limitations in hybrid cell generation.

Purpose of the Study:

• To develop and validate a novel, highly efficient single-cell fusion system for cellular reprogramming.
• To demonstrate the successful reprogramming of porcine somatic cells and pluripotent stem cells using this new method.

Main Methods:

• Development of a single-cell fusion technique, optimized for improved efficiency.
• Fusion experiments involving mouse embryonic stem cells (mESCs) and porcine cells (PEFs and pPSCs).
• Verification of hybrid cell formation through fluorescence and species-specific gene expression analysis.

Main Results:

• The single-cell fusion system demonstrated significantly higher efficiency than traditional PEG fusion.
• Successfully generated hybrid cells expressing both mouse and porcine genes, confirmed by fluorescence and molecular markers.
• Generated hybrid cells exhibited pluripotency characteristics similar to mESCs, including colony morphology and embryoid body formation.
• Key pluripotency factors (OCT4, NANOG, SOX2) were induced in porcine cells within the hybrid cells.
• The transforming growth factor-beta signaling pathway was implicated in maintaining porcine pluripotency.

Conclusions:

• The single-cell fusion method is highly efficient for cellular reprogramming.
• This technique enables the reprogramming of porcine embryonic fibroblasts and pluripotent stem cells by mouse embryonic stem cells.
• The generated hybrid cells offer a valuable tool for investigating porcine pluripotency and signaling pathways, with potential applications in regenerative medicine.