The invention involves improving the cultivation of T cells by incubating them with short-chain fatty acid (SCFA) pentanoate after isolation from peripheral blood. The effect is that the cells are activated and the production of effector molecules is increased. This increases the chances of success of tumor therapy. This is illustrated by T-cells from mice that are transferred to mice with subcutaneous pancreatic tumors after the procedure. This type of cell treatment can be transferred to humans and the improved treatment of pancreatic cancer. We show in detail that the short-chain fatty acid (SCFA) pentanoate enhances the function of CD8+ cytotoxic T lymphocytes (CTLs). We show that Pentanoate promotes the core molecular signature of murine CD8+ CTLs. Pentanoate enhances anti-tumor activity of antigen-specific CTLs. Bacterial-derived SCFAs exhibit specific HDAC class I inhibitory activity. Pentanoate-producing bacteria enhance CD8+ T cell-mediated anti-tumor immune responses.

One aspect of the present disclosure can include a priming medium for creating an isolated population of stem cells having an anti-inflammatory phenotype from an unprimed population of stem cells. The priming media can include a serum-free medium, a functional activator of a Type I interferon (IFN) pathway and a Type II IFN pathway, and at least two pro-inflammatory cytokines. The functional activator and the at least two pro-inflammatory cytokines can be present in an amount sufficient to promote induction of stem cells having an anti-inflammatory phenotype. The cells having an anti-inflammatory phenotype can be marked by increased expression and/or secretion of one or more anti-inflammatory or immune modulatory mediators as compared to the unprimed population of stem cells. Other aspects of the present disclosure can include stem cells made according to the present disclosure as well as therapeutic compositions and uses of the stem cells.

The present invention relates to a chemically defined medium for eukaryotic cell culture, comprising water, at least one carbon source, one or more vitamins, one or more salts, one or more growth factors, one or more fatty acids, one or more buffer components, selenium and one or more further trace elements and its use in the culture of cancer stem cells, in particular tumorsphere culture of cancer stem cells.

The present invention relates in part to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed.

The present invention provides a new method for producing Engineered Heart Muscle (EHM) under chemically fully defined conditions all compatible with GMP regulations. The resulting human myocardium generates force and shows typical heart muscle properties.

A transfection reagent composition comprising: 30-60 MOL. % of a cationic lipid, or a pharmaceutical acceptable salt thereof; 10-60 MOL % of a structural lipid; 10-20 MOL % of a triglyceride; and 0.1 to about 10 MOL. % of a stabilizing agent, is provided. The transfection agent is effective in transfecting cells, particularly neurons, with siRNA, mRNA and plasmid nucleic acid, Sand maintaining viability of the cells as well as activity of the delivered nucleic acid.

The disclosure provides methods and compositions for affecting the development of antigen presenting cell (APC, e.g., a macrophage or dendritic cell). The methods include maturing an APC, promoting anti-inflammatory phenotype, promoting development of a T regulatory cell (Treg) from a naive T cell. The methods generally include exposing an APC to a tryptophan derived microbiota metabolite (TDMM), such as an anti-inflammatory or pro-mucosal TDMM, and permitting the APC to mature. In some embodiments, the conditioned APC is exposed to a naive T cell to further promote development of a T regulatory cell (Treg). In some embodiments, the TDMM is selected from the group consisting of indole, indole-3-acetate, 5-hydroxyindole, and indole-3-pyruvate.

The present invention discloses formulation of a serum-free medium used for human pluripotent stem cells, which comprises the following raw materials: inorganic salt components, organic components, amino acids and amino acid salts, energy substances and metabolic intermediates, vitamins and antioxidants, proteins and polypeptides, trace elements and chromogenic substances; while the culture process comprises the following steps: selecting a basic formulation, performing combination screening, identifying and evaluating results, and testing a new formulation of culture; and proportioning according to the following methods: adding aforesaid raw materials into 950 ml of water for injection, stirring gently until dissolved, and finally adding 2.438 g of sodium bicarbonate, and stirring gently until dissolved, and then adding 1 liter of water for injection, adjusting the pH to the desired value with 1 mol/L sodium hydroxide solution or 1 mol/L hydrochloric acid solution, finally filtering sterilized with 0.1 μm diameter filter under positive pressure, and storing the medium solution in dark place at 2° C.-8° C., the invention solves the problem of high cost of domestic import of serum-free formulation.

Provided are novel serum-free culture media which comprise basic fibroblast growth factor (bFGF), transforming growth factor beta-3 and ascorbic acid at a concentration of at least about 50 microgram/ml; ascorbic acid at a concentration range of about 400-600 microgram/ml, bFGF at a concentration range of about 50-200 ng/ml, xeno-free serum replacement and a lipid mixture; the IL6RIL6 chimera at a concentration range of about 50-200 picogram per milliliter (pg/ml); or leukemia inhibitory factor (LIF) at a concentration of at least 2000 units/ml; cell cultures comprising same with pluripotent stem cells such as human embryonic stem cells and induced pluripotent stem (iPS) cells, and methods of using same for expanding pluripotent stem cells in an undifferentiated state using two-dimensional or three-dimensional culture systems; and methods of expanding iPS cells in a suspension culture devoid of substrate adherence and cell encapsulation.

The methods and compositions described herein surprisingly increase CRISPR/Cas-mediated gene editing in stem cells by transiently treating the cells with a stem cell viability enhancer prior to and/or after contacting the cells with one or more CRISPR/Cas9 components. Further, this treatment also surprisingly results in increased engraftment of the stem cells into the target tissue of a subject. The present disclosure also provides one or more modified CRISPR/Cas9 components which, when used in combination with the stem cell viability enhancer, further increases the frequency of gene editing in stem cells, increases stem cell viability, and increases stem cell engraftment.