Blastocyst complementation generates exogenous donor-derived liver in ahepatic pigs

Area of Science:

• Xenotransplantation
• Organogenesis
• Developmental Biology

Background:

• Liver diseases cause significant global morbidity and mortality, with limited donor organs for transplantation.
• Current organ transplantation faces challenges due to stringent criteria and organ scarcity.
• Generating chimeric organs in pigs is a potential solution to bridge the organ shortfall.

Purpose of the Study:

• To investigate the feasibility of generating chimeric livers in pigs.
• To utilize a conditional blastocyst complementation approach for organogenesis.
• To enable organ development using donor-derived pluripotent cells in a vacant niche.

Main Methods:

• Sequential genetic modification of porcine fetal fibroblasts (FOXA3^CREHHEX^loxP/loxP).
• Somatic cell nuclear transfer using a conditional HHEX knockout and GFP donor (COL1A^{CAG:LACZ 2A EGFP}).
• Generation of host embryos, complementation, and transfer into surrogate mothers.

Main Results:

• Successful generation of chimeric pig fetuses.
• Donor EGFP blastomeres were confirmed to reconstitute hepatocytes via immunohistochemistry.
• Demonstrated feasibility of exogenous donor-derived hepatogenesis in a large animal model.

Conclusions:

• The study demonstrates a viable approach for creating chimeric livers in pigs.
• Conditional blastocyst complementation enables donor cell contribution to organ development.
• This research paves the way for future advancements in regenerative medicine and organ transplantation.