Breeding and identification of Wuzhishan miniature pigs with α-1, 3 galactosyltransferase gene-knockout

  Objective  To summarize the breeding and identification of Wuzhishan miniature pig models with α-1, 3 galactosyltransferase (GGTA1) gene-knockout (GTKO).

  Methods  The breeding and reproduction perform of GTKO Wuzhishan miniature pigs were assessed and the quantity of piglets was counted. The GTKO Wuzhishan miniature pig models with GGTA1gene knockout were validated by polymerase chain reaction (PCR). The αGal phenotype of peripheral blood mononuclear cells (PBMC) in human, wild-type Wuzhishan miniature pigs and GTKO Wuzhishan miniature pigs was detected by fluorescent microscope and flow cytometry. Routine blood test parameters were statistically compared between the GTKO and wild-type Wuzhishan miniature pigs.

  Results  The inheritance of GGTA1 genotype complied with Mendel's law. Flow cytometry detected no fluorescent expression of PBMC in GGTA1^－/－ pig models, which were consistent with the genotype identification results. The mean piglets of the primiparous GTKO Wuzhishan miniature pigs were (6.8±1.8) and (8.3±2.2) for the multiparous Wuzhishan miniature pigs. No statistical significance was noted in routine blood test parameters between the GTKO and wild-type Wuzhishan miniature pigs (all P>0.05).

  Conclusions  Stable inheritance and normal reproductive capacity are observed in two generations of Wuzhishan miniature pigs continuously. GTKO Wuzhishan miniature pig is a reliable donor for heterogeneous organ transplantation.