Selection of genetically modified chicken blastodermal cells by magnetic-activated cell sorting

Area of Science:

• Animal Biotechnology
• Developmental Biology
• Cell Biology

Background:

• Production of transgenic chickens relies on genetic modification of chicken blastodermal cells (CBC).
• Higher proportions of stably transfected CBC in embryo inoculum correlate with increased chimeric bird frequency.
• Enrichment of transfected CBC is crucial for efficient generation of transgenic poultry.

Purpose of the Study:

• To evaluate magnetic-activated cell sorting (MACS) as a method for enriching transfected CBC.
• To assess the effectiveness of MACS in isolating genetically modified chicken cells.
• To determine if magnetically sorted CBC retain their developmental potential.

Main Methods:

• Chicken blastodermal cells were coelectroporated with pmiwZ and pMACS Kk plasmids.
• Cells were sorted using magnetic-activated cell sorting (MACS) based on mouse H-2Kk surface molecule expression.
• Transfection efficiency was assessed by X-gal staining for lacZ expression.

Main Results:

• MACS resulted in a 1.4-fold enrichment of lacZ-positive CBC after 48 hours of culture.
• The enriched cell population contributed to both extra-embryonic and intra-embryonic tissues in recipient embryos.
• A marginal increase in intra-embryonic contribution was observed from the magnetically sorted cells.

Conclusions:

• Magnetic-activated cell sorting (MACS) effectively enriches genetically modified chicken blastodermal cells (CBC).
• Magnetically sorted CBC maintain their developmental potential, contributing to ectodermal and mesodermal lineages.
• This technique shows promise for enhancing the production of transgenic birds through improved genetic modification efficiency.