The present invention provides methods and compositions for making proteins, preferably antibodies, more preferably anti-tumor necrosis factor alpha antibodies, and most preferably adalimumab. The present invention further provides methods and compositions for mammalian cell culture, preferably Chinese Hamster Ovary cells.

Provided herein are enriched populations of ventricular compact cardiomyocytes and enriched populations of mature ventricular or atrial cardiomyocytes, as well as methods of generating the enriched cell populations and methods of using the enriched cell populations in regenerative cardiac cell therapies.

The present invention relates to a biologically active placenta-derived liquid human substrate (hpS) comprising extracellular matrix (ECM) proteins, cytokines and growth factors and use thereof. The present invention also provides methods of producing a composition comprising biologically active placenta-derived liquid human substrate (hpS).

A method of producing Klotho protein includes preparing a Klotho plasmid DNA vector, culturing cells, transfecting the cells with the Klotho plasmid DNA vector in a cell culture medium, growing the transfected cells, and harvesting the cell culture supernatant by removing the transfected cells. The Klotho plasmid DNA vector has a mammalian selection marker and a Klotho open reading frame. The cells are primary fibroblast cells and/or mesenchymal stromal cells. A method of manufacturing a cosmetic composition includes combining Klotho protein or the cell culture supernatant with a cosmetically acceptable vehicle. A method of treating a patient to improve the condition and appearance of aging skin includes topically administering the cosmetic composition to the patient. By upregulating the Klotho gene in vitro and incorporating the Klotho protein and growth factors into a composition, transepidermal water loss, skin atrophy, and free radical damage to the skin may be addressed.

The present invention relates to a cell culture medium for preparing liver, gastric, pancreatic, colon or intestinal adult stem cell isolated from adult tissue, as well as for maintaining such stem cell in the undifferentiated state. The cell culture medium comprises a base medium; an ABL and SRC dual kinase inhibitor/an ABL kinase inhibitor and a SRC kinase inhibitor; a mitogenic factor; a WNT signalling pathway activator; a stimulator for NAD+ and NADP+ generation; and a cAMP/P KA pathway activator. In a particular embodiment, the ABL and SRC dual kinase inhibitor is Dasatinib; the mitogenic factor is EGF; the WNT signalling pathway activator is R-Spondin 1; the stimulator for NAD+ and NADP+ generation is nicotinamide; and the cAMP/PKA pathway activator is cholera endotoxin.

Hematopoeitic stem/progenitor cells (HSPC) and/or non-T effector cells are modified to express an extracellular component including a tag cassette. The tag cassette can be used to activate, promote proliferation of, detect, enrich, isolate, track, deplete and/or eliminate modified cells. The cells can also be modified to express a binding domain.

A chemically defined medium for differentiation of muscle stem cells in vitro, namely a serum-free, more efficient and inexpensive chemically defined medium for inducing differentiation of muscle stem cells in vitro. Compared with an existing general muscle stem cell differentiation medium, using the chemically defined medium can increase the relative expression of myogenin genes by 4.48 times on the 2.sup.nd day of differentiation, increase the relative expression of myosin heavy chain genes by 55.28 times on the 6.sup.th day, and increase the percentage of cell differentiation from 34.94% to 57.93% in the terminal differentiation stage, and more, thicker and longer muscle fibers are formed through induced differentiation. The chemically defined differentiation medium further improves the differentiation efficiency of muscle stem cells, and provides a more efficient and inexpensive method for differentiation of muscle stem cells into myotubes, and for 3D culture of muscle stem cells to produce cell cultured meat.

A method for generating a three-dimensional neuromuscular organoid in vitro is disclosed. This method comprises the following steps: a) providing a first cell culture comprising neuromesodermal progenitor cells and cultivating the neuromesodermal progenitor cells in a first differentiation medium chosen from the group consisting of i) a non-supplemented serum-free cell culture medium and ii) a serum-free cell culture medium supplemented with at least one of a ROCK inhibitor, an activator of a growth factor signaling pathway, and an activator of an insulin signaling pathway; b) replacing the first differentiation medium by a second differentiation medium within 1 to 3 days after cultivation start, wherein the second differentiation medium is chosen from the group consisting of i) a non-supplemented serum-free cell culture medium and ii) a serum-free cell culture medium supplemented with at least one of an activator of a growth factor signaling pathway, and an activator of an insulin signaling pathway; c) replacing the second differentiation medium by a non-supplemented serum-free cell culture medium within 1 to 3 days after replacing the first differentiation medium by the second differentiation medium; and d) obtaining a three-dimensional neuromuscular organoid from the non-supplemented serum-free cell culture medium.

Method and compositions for inducing the self-renewal of stem/progenitor supporting cells comprised by a cochlear cell population, including inducing the stem/progenitor cells to proliferate while maintaining, in the daughter cells, the capacity to differentiate into hair cells.

Disclosed are means, methods and compositions of matter useful for inhibiting, in an antigen-specific manner, immunity towards an autoantigen or alloantigen. In one embodiment of the invention, regenerative cells are cultured ex vivo together with immune cells from a mammal suffering from an autoimmune condition. Autoantigens or alloantigens are added in the culture of regenerative cells and cells from an autoimmune disease suffering individual in a manner so that said regenerative cells can endow onto said immune cells of said patient suffering from autoimmunity a state of antigen specific infectious tolerance. In one embodiment, said infectious tolerance involves T regulatory cells inducing conversion of dendritic cells to tolerogeneic dendritic cells, and furthermore in other embodiments administration of tolerogenic dendritic cells induces T regulatory cells.