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AssigneesBresaGen Limited, an Australia corporation, St. Vincent's Hospital and Anthony J. D'Apice of Australia
InventorsAnthony J.F. d'Apice, Martin J. Pearse, Allan J. Robins, Robert J. Crawford, Peter D. Rathjen
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Human pre-formed xenoantibodies play an important role in the hyperacute rejection response in human xenotransplantation. Disclosed are materials and methods for removing or neutralizing such antibodies. Also disclosed are materials and methods for reducing or eliminating the epitopes in the donor organs that are recognized by such antibodies. Such epitopes are formed as the result of activity by the enzyme α-1,3 galactosyltransferase. The porcine gene encoding α-1,3 galactosyltransferase is disclosed, as are materials and methods for inactivating (“knocking out”) the α-1,3 galactosyltransferase gene in mammalian cells and embryos. Included are nucleic acid constructs useful for inactivating the α-1,3 galactosyltransferase gene in a target cell. Also disclosed is a novel leukemia inhibitory factor (T-LIF) that is useful for maintenance of embryonic stem cells and primordial germ cells in culture.
A method for generating a porcine cell comprising at least one inactivated α-1,3 galactosyltransferase gene, the method comprising:
A porcine cell comprising at least one disrupted α-1,3 galactosyltransferase gene, wherein the disruption is by insertion of an exogenous sequence into said gene such that the disruption prevents expression of functional α-1,3 galactosyltransferase and wherein the gene, prior to disruption, encodes the porcine α-1,3 galactosyltransferase with an amino acid sequence of SEQ ID NO:10, wherein the cell is in vitro.
A method for eliminating or reducing hyperacute rejection of non-primate mammalian cells, tissues and organs by human serum, the method comprising adding, to said human serum, a physiologically acceptable amount of galactose or a saccharide in which the terminal carbohydrate is an a galactose linked at position 1, prior to exposure of said-human serum to said non-primate cells, wherein said amount of galactose or saccharide is sufficient to reduce or eliminate said hyperacute rejection.
Dependencies are inferred from the claim text.