Transgenesis and nuclear transfer using porcine embryonic germ cells

Area of Science:

• Reproductive Biology
• Stem Cell Research
• Genetics

Background:

• Embryonic germ (EG) cells are derived from primordial germ cells (PGCs) and possess indefinite proliferation potential.
• EG cells can differentiate both in vitro and in vivo, making them promising for various applications.
• Their long-term culture stability suggests suitability as nuclear donor cells for nuclear transfer (NT).

Purpose of the Study:

• To compare the efficiency of transgenesis and NT using porcine EG cells versus fetal fibroblasts.
• To evaluate the potential of EG cells as a source for generating transgenic pigs.
• To assess the developmental competence of NT embryos derived from EG cells.

Main Methods:

• Isolation and culture of porcine EG cells and fetal fibroblasts.
• Introduction of the green fluorescent protein (GFP) gene into porcine EG cells.
• Porcine nuclear transfer (NT) using EG cells and fetal fibroblasts as donor cells.
• Assessment of blastocyst development rates and GFP expression in NT embryos.

Main Results:

• EG cell NT resulted in a significantly higher rate of blastocyst development (18.2%) compared to somatic cell NT (14.4%).
• Nuclear transfer embryos derived from GFP-transfected EG cells successfully developed into blastocysts expressing GFP (21.2%).

Conclusions:

• Porcine EG cell NT demonstrates superior developmental rates compared to somatic cell NT.
• EG cells are a viable source for producing transgenic pigs through nuclear transfer and gene introduction.