The present invention provides tissues derived from animals, which lack any expression of functional alpha 1,3 galactosyltransferase (alpha-1,3-GT). Such tissues can be used in the field of xenotransplantation, such as orthopedic reconstruction and repair, skin repair and internal tissue repair or as medical devices.

The invention provides for compositions and methods for making and using adipose-derived stem cells for treating non-human mammals for various medical conditions. In particular, the invention provides methods and compositions useful for repair of bone fractures, and for treatment of dry eye conditions, acute renal failure, and chronic renal failure in non-human mammals. The ADSC are porcine adipose tissue-derived stem cells (ADSC) isolated from transgenic porcine animals.

Disclosed herein, are inducible immunodeficient animals and methods to make them by adding an IL2Rg/RAG2 rescue cassette (RG-reg) or an IL2Rg/RAG2/FAH rescue cassette (FRG-reg) to a line of IL2Rg/RAG2 knockout (RG-KO) or IL2Rg/RAG2/FAH knockout (FRG-KO) swine. The rescue cassette enables line breeding of immunocompetent (regRG-KO) or (regFRG-KO) swine for rapid propagation. The rescue cassette can be excised, specifically in germ cells of regRG-KO or regFRG-KO swine, such that offspring of animals do not possess the rescue cassette and are immunodeficient. The immunodeficient swine also provide host embryos having genetic ablations to provide a niche for organ complementation by human stem cells.

The present invention relates to a transgenic pig in which an immune rejection response is suppressed during xenotransplantation, wherein a gene coding for heme oxygenase-1 (HO-1) and a gene coding for tumor necrosis factor receptor 1-Fc (TNFR1-Fc) are simultaneously expressed and a gene coding for -1,3-galactosyltransferase (GGTA1) is knocked out; and a method for producing the same.

The transgenic pig of the present invention, in which the genes coding for human HO-1 and TNFR1-Fc fusion protein are simultaneously expressed and the gene coding for GGTA1 is knocked out, may reduce oxidative stress during organ isolation and in vitro culture by antioxidative reaction, cytoprotective function, etc., and may also reduce a TNF--mediated inflammatory response in early transplantation by TNFR1-Fc expression. In addition, the transgenic pig may inhibit the maturation of dendritic cells and regulate the activation and proliferation of T-cells, thereby reducing an acute vascular rejection response to promote early engraftment of a transplanted organ. In addition, the transgenic pig can increase the viability of a transplanted organ by suppressing a hyper-acute immune rejection reaction caused by GGTA1. Accordingly, an organ, in which an immune rejection response is suppressed during xenotransplantation, can be produced using the transgenic pig.

In human-human transplants, the organ recipient's serum is tested against a broad array of HLA (human leukocyte antigens) alleles. The current clinical assay almost performs a virtual crossmatch which allows elimination of incompatible donor organs in advance, but the current assay presents fragments of HLA polypeptides that are not normally visible to antibodies. As a result, the assay yields false positives. Further the current clinical assay is optimized for human to human transplant, not swine to human transplant. Some HLA antibodies also bind swine leukocyte antigens (SLA). Rather than using beads to present antigens, the compositions and methods provide cellular presentation of potential antigens. The application provides a modified human C1R cell line with significantly reduced antigenicity to human sera. Multiple cell lines, each expressing a different SLA, were created. The modified C1R line may express HLAs or other potential antigens of interest. The application also provides modified HEK293T cells for antigen display.

Disclosed are genetically modified porcine cells, tissues, organoids, and animals and methods of using the same for transplantation, and testing for xenoreactivity.

To provide a method of efficiently producing full thickness skin having skin accessory organs. [Solution] This method of producing full thickness skin having skin accessory organs is characterized by involving the following steps (a) to (d): (a) a step for stimulating an embryoid body with a physiologically active substance that can activate the Wnt pathway, (b) a step for preparing a conjugate that includes all or part of the embryoid body stimulated in step (a) and a scaffolding material, (c) a step for transplanting the conjugate prepared in step (b) into an animal, and (d) a step for producing, in the animal, full thickness skin derived from said conjugate.

A revised technique for preparing human tissues and organs aiming at application to medical use is provided. With this technique, function per cell is greatly improved and great reduction in production cost is realized.

A method of preparing an organ bud, comprising culturing vascular endothelial cells, mesenchymal cells and a tissue or organ cell in vitro in the presence of blood cells. Also provided are an organ bud prepared by the above method and a method of preparing a tissue or an organ using the organ bud. Using the same organ bud, there are provided a method of transplanting an organ bud, a method of regeneration or function recovery of a tissue or an organ, a method of preparing a non-human chimeric animal, and a method of evaluating a drug.

The present invention is a porcine animal, tissue, organ, cells and cell lines, which lack any expression of functional alpha 1,3 galactosyltransferase (alpha1,3GT). These animals, tissues, organs and cells can be used in xenotransplantation and for other medical purposes.

A method of modulating some or all copies of a gene in a cell is provided including introducing into a cell one or more ribonucleic acid (RNA) sequences that comprise a portion that is complementary to all or a portion of each of the one or more target nucleic acid sequences, and a nucleic acid sequence that encodes a Cas protein and maintaining the cells under conditions in which the Cas protein is expressed and the Cas protein binds and modulates the one or more target nucleic acid sequences in the cell.