The invention relates to a method for screening an optimal
gene combination of a heterograft donor pig and belongs to the technical field of animal biologics. By taking human
fibroblast as a study target, an in-vitro compatibility evaluation method for human
fibroblast is established, results of reported pig
fibroblast modified with heart or other genes with long
transplantation survival times canbe verified by using the method, and relatively applicable complement adjustment factors, immune response factors and anticoagulation factors can be screened; on the basis of GTKO (
Gene Transferase Knockout), the factors are combined and are subjected to fixed knock-in or knockout, pig fibroblast of different multi-factor combinations is subjected to early-stage screening, then cells of more thanthree optimal
gene combinations are recloned, cloned pigs are subjected to
heart transplantation of pig-non-
primate animals, and the reliability of the
screening method can be verified. By adopting the method, an optimal
gene combination can be evaluated without the basis that a gene combination modified pig is available, so that the gene combination screening time and the
investment cost can begreatly reduced, and the method has great significances in development of real heterograft donor pigs.