Targeting αGal epitopes for multi-species embryo immunosurgery

Area of Science:

• Reproductive biology
• Immunology
• Xenotransplantation

Background:

• Inner cell mass (ICM) isolation relies on complement-mediated lysis of trophectoderm (TE) cells.
• Conventional methods use undefined antibodies, limiting specificity and applicability.
• Genetically engineered pigs (GTKO) lack alpha-1,3-galactose (αGal) but produce anti-αGal antibodies due to exposure.

Purpose of the Study:

• To investigate the potential of GTKO pig serum as a novel antibody source for multi-species embryo immunosurgery.
• To determine if anti-αGal antibodies in GTKO pig serum can lyse TE cells across different species.
• To assess the efficacy of GTKO pig serum for isolating pure intact ICMs.

Main Methods:

• Utilized mouse, rabbit, pig, and cattle blastocysts.
• Detected αGal epitope expression on TE cells of tested species.
• Applied GTKO pig serum for complement-mediated lysis of TE cells.
• Evaluated the purity and integrity of recovered ICMs.

Main Results:

• αGal epitopes were present on TE cells of all tested species.
• GTKO pig serum induced effective complement-mediated lysis of TE cells.
• Intact ICMs were successfully isolated with high purity across species.
• Demonstrated the efficacy of GTKO pig serum in multi-species embryo immunosurgery.

Conclusions:

• GTKO pig serum is a reliable and effective source of antibodies for targeting αGal epitopes on TE cells.
• This approach facilitates multi-species embryo immunosurgery, enhancing research capabilities.
• Offers a novel, species-independent method for isolating pure ICMs.