The invention provides methods of preparing a sample of viable diseased cells obtained from a human subject for clinical testing, wherein the methods inhibit anoikis and/or anoikis in the cells while maintaining the physiological functions and genomic composition of the cells when they were in vivo. In the methods of the invention, primary cells are cultured in media comprising at least one anoikis inhibitor, preferably at least one inhibitor of an intrinsic anoikis pathway and at least one inhibitor of an extrinsic anoikis pathway, under anti-anoikis atmospheric conditions, such as greater than 2% and less than 20% oxygen. Method combining multiple culturing conditions, including surface attachment under conditions that inhibit anoikis, are also provided. Compositions and kits for use in the methods of the invention are also provided.

The present invention is directed to compositions and methods to increase the expression of PD-L1 and/or IDO-1 in a population of cells, the modulated cells expressing increased PD-L1 and/or IDO-1, and methods related to the immunosuppressive effects obtained by cells expressing increased PD-L1 and/or IDO-1.

The invention provides methods of preparing a sample of viable diseased cells obtained from a human subject for clinical testing, wherein the methods inhibit anoikis and/or anoikis in the cells while maintaining the physiological functions and genomic composition of the cells when they were in vivo. In the methods of the invention, primary cells are cultured in media comprising at least one anoikis inhibitor, preferably at least one inhibitor of an intrinsic anoikis pathway and at least one inhibitor of an extrinsic anoikis pathway, under anti-anoikis atmospheric conditions, such as greater than 2% and less than 20% oxygen. Method combining multiple culturing conditions, including surface attachment under conditions that inhibit anoikis, are also provided. Compositions and kits for use in the methods of the invention are also provided.

According to the present disclosure, provided is a method for manufacturing induced pluripotent stem cells including preparing cells and introducing RNA into the cells, wherein the RNA includes RNA encoding a reprogramming factor and wherein, in the RNA introduced into the cells, double-stranded RNA is substantially removed.

Methods of treating T cells with Venetoclax to increase T cell-mediated cytotoxicity and/or T cell mediated anti-tumor activity are described. Also described are populations of enhanced T cells as well as associated methods and uses for the treatment of cancer.

The present specification provides methods for augmenting the antigenic content, especially of tumor-associated antigens (TAA), and immunogenicity of cancer cells; methods for enhancing cross-presentation in dendritic cells, compositions comprising such manipulated cells derived from single cancer patients; and methods of using those compositions as a personal immunotherapeutic product to treat the donor patient's cancer.

The present disclosure is broadly concerned with the field of cancer immunotherapy. For example, the present invention generally relates to an immune cell comprising a genetically engineered antigen receptor that specifically binds to a target antigen and a genetic disruption agent that reduces or is capable of reducing the expression in the immune cell of two genes that weaken the function of the immune cell.

Glycoproteins that are transgenically produced in mammalian cells exhibit non-human glycan structures. As in humans, this can possibly lead to immune responses, the drug manufacturing potential of these drugs is limited. On the other hand, recombinant protein production in human cells is inefficient due to the cells' poor protein yields, proliferation potential and cellular density. The present application solves these issues by providing a recombinant vertebrate cell that is comprising a non-vertebrate and/or artificial phosphatidylethanolamine-binding protein (PEBP). Compared to a parent cell line, the recombinant cells of the invention exhibit improved cell growth, protein yield and excellent compatibility with other established protein production methods. Furthermore, methods, for producing a cell line with improved vitality, protein expression and cell growth characteristics by introducing a non-vertebrate and/or artificial PEBP is given. Moreover, both a nucleic acid construct that is suitable for regulating recombinant protein expression in a cell by coding for such a PEBP and a recombinant cell comprising such a nucleic acid construct is provided. Lastly, a method for the recombinant expression of a target protein by culturing such a recombinant vertebrate cell of the invention is given, wherein the cell is also comprising an expression construct encoding the target protein.

A method to enhance homology directed recombination (HDR) efficiency, and a kit therefor, are provided.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating said genetic modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes a survival factor, wherein the genetic modification comprises an insertion of a polynucleotide encoding a tolerogenic factor. The universal donor cells may further comprise at least one genetic modification within or near a gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or a component or a transcriptional regulator of a MHC-I or MHC-II complex, wherein said genetic modification comprises an insertion of a polynucleotide encoding a second tolerogenic factor.