The present disclosure relates to the field of cell therapy, and more specifically, to improving CAR and/or TCR function through improvement of the tumor microenvironment via improvement in cytokine signaling.

The invention provides methods for using Umbilical Cord Lining Stem Cells (ULSCs) to produce therapeutic factors including growth factors, cytokines, chemokines and extracellular matrix components. ULSCs are mesenchymal stem cells isolated from umbilical cord lining. They can be efficiently propagated and expanded in vitro. Under specific conditions ULSCs produce useful therapeutic factors that can be used to treat injuries and degenerative conditions.

Factor rich compositions produced from umbilical cord (UC) mesenchymal stem cells (MSCs) are described. Secretory UC MSCs in serum free culture conditions produce a factor rich conditioned medium which may be concentrated and filtered to obtain clinical grade products.

Compounds that either produced a higher proportion or greater absolute number of phenotypically identified nave, stem cell memory, central memory T cells, adaptive NK cells, and type I NKT cells are identified. Compositions and methods for modulating immune cells including T, NK, and NKT cells for adoptive cell therapies with improved efficacy are provided.

A composition for inducing and/or amplifying T.sub.SCM in vitro, a culture medium including the composition, and a method for inducing and/or amplifying T.sub.SCM in vitro are provided, wherein the composition comprises inducing agents including IL-7 and IL-21. The chimeric antigen receptor T-memory stem cells induced differentiated and amplified by adding the composition can be used directly for reinfusion therapy of patients.

A method of culturing natural killer (NK) cells according to an embodiment of the present disclosure can be used in immunotherapy. An additive composition for an NK cell culture medium according to an embodiment of the present disclosure may inhibit the activity of CD4+ T cells, thereby effectively helping NK cell proliferation and activation.

The present invention relates to: a method for producing immunocytes, specifically induced natural killer T (iNKT) cells that are induced by direct reprogramming of isolated somatic cells, and chimeric antigen receptor (CAR)-iNKT cells into which a CAR gene encoding a CAR is introduced; iNKT cells produced by the method; and a cell therapy composition and a pharmaceutical composition for preventing or treating cancer, comprising the iNKT cells.

The method according to the present invention can produce, through direct reprogramming, iNKT cells or iNKT cells into which a CAR gene is introduced, from isolated cells so as to simplify the production process and shorten production time, thereby reducing costs, to have excellent NKT cell production efficiency, and to ensure safety according to the production without passing through induced pluripotent stem cells, thereby having an excellent NKT cell production effect distinguished from that of a conventional reprogramming technique. In addition, the iNKT cells or iNKT cells into which a CAR gene is introduced, which are produced by the method, have an excellent cancer cell killing ability, and thus can be effectively used as a cell therapy composition or a pharmaceutical composition for preventing or treating cancer.

Cell culture media, preservative media or cryopreservation media include a low dose of one or more cytokines, e.g. interleukin-2 (IL-2).

The disclosure relates to methods of culturing and expanding CD4+ and/or CD8+ T cells in culture. In some embodiments, the methods include expanding, proliferating and storing lymphocytes in tissue culture by exposing the lymphocytes to a combination of cytokines and/or nucleic acids expressing cytokines or functional fragments or variants thereof. The disclosure further relates to methods of generating and manufacturing CD4+ and/or CD8+ T cells that are specific to one or a plurality of viral antigens.

The present disclosure relates to methods that include the use of a first multi-chain chimeric polypeptide and a second multi-chain chimeric polypeptide for stimulating the NK cells, inducing or increasing proliferation of the NK cells, inducing differentiation of the NK cells, and treating a subject in need thereof using activated NK cells.