Provided are a method for preparing genetically engineered erythrocytes carrying an anti-PD-1 single chain antibody, and genetically engineered erythrocytes carrying the anti-PD-1 single chain antibody. The preparation method comprises the following steps: constructing a desired fragment sequence in a lentiviral expression vector, lentivirally packaging the vector of the desired sequence to obtain a high-titer lentiviral concentrate; isolating Lin.sup.?CD34.sup.? cells from peripheral blood mononuclear cells and enriching the Lin.sup.?CD34.sup.? cells; inducing the Lin.sup.?CD34.sup.? cells to differentiate into erythroid and performing proliferation thereon; using the lentiviral concentrate to infect the Lin.sup.?CD34.sup.? cells; and obtaining mature anti-PD-1 scFv erythrocytes via erythrocyte denucleation. The present genetically engineered erythrocytes carrying the anti-PD-1 single chain antibody can perform the targeted delivery of an anti-PD-1 single-chain antibody to tumor tissues.

Disclosed herein are bioreactor systems and methods of utilizing said systems.

Provided herein are methods for identifying and treating cancers that are resistant to PARP inhibition. Methods for sensitizing cancers to a PARP inhibitor therapy are also provided. In some aspects. PARP inhibitor cancers are treated with a PARP inhibitor therapy in combination with a c-Met inhibitor therapy.

There is described an isolated 3-dimensional liver spheroid wherein said spheroid has: increased ATP content as compared to a 3-dimensional liver spheroid cultured in Complete William's E medium alone; the same or increased activity of cytochrome P450 1A1 and cytochrome P450 1B1 as compared to a 3-dimensional liver spheroid cultured in Complete William's E medium alone; and increased albumin secretion as compared to a 3-dimensional liver spheroid cultured in William's E medium alone.

Disclosed are means, methods, and compositions of matter useful for augmentation of a vaccine induced antitumor immune response through immunological targeting of tumor endothelium. In one embodiment a cancer antigen specific vaccine is combined with an endothelial targeting vaccine in a manner to facilitate release of tumor antigens as a result of endothelial destruction. The released tumor antigens serve to amplify immune response induced by the cancer antigen specific vaccine. In one embodiment an endogenous cancer vaccine is utilized as a source of immunization. The endogenous cancer vaccine may be cancer cells induced to die locally such as by means of: hyperthermia, irradiation, oncolytic virus exposure, and embolization.

A method for producing an epithelial cell sheet, comprising culturing cells derived from oral mucosal epithelial cells on a substrate in a serum-free medium, wherein the serum-free medium comprises (i) EGF protein or KGF protein, (ii) B-27 supplement, and (iii) a ROCK inhibitor.

An in vitro microfluidic intestine on-chip is described herein that mimics the structure and at least one function of specific areas of the gastrointestinal system in vivo. In particular, a multicellular, layered, microfluidic intestinal cell culture, which is some embodiments is derived from patient's enteroids-derived cells, is described comprising L cells, allowing for interactions between L cells and gastrointestinal epithelial cells, endothelial cells and immune cells. This in vitro microfluidic system can be used for modeling inflammatory gastrointestinal autoimmune tissue, e.g., diabetes, obesity, intestinal insufficiency and other inflammatory gastrointestinal disorders. These multicellular-layered microfluidic intestine on-chips further allow for comparisons between types of gastrointestinal tissues, e.g., small intestinal duodenum, small intestinal jejunum, small intestinal ileum, large intestinal colon, etc., and between disease states of gastrointestinal tissue, i.e. healthy, pre-disease and diseased areas. Additionally, these microfluidic gut-on-chips allow identification of cells and cellular derived factors driving disease states and drug testing for reducing inflammation.

There is described an isolated 3-dimensional liver spheroid wherein said spheroid has: increased ATP content as compared to a 3-dimensional liver spheroid cultured in Complete William's E medium alone; the same or increased activity of cytochrome P450 1A1 and cytochrome P450 1B1 as compared to a 3-dimensional liver spheroid cultured in Complete William's E medium alone; and increased albumin secretion as compared to a 3-dimensional liver spheroid cultured in William's E medium alone.

Provided is a method for producing an economical bovine serum composition containing many factors useful for cell proliferation. The method includes a step of performing an anticoagulation treatment of bovine whole blood with an anticoagulant, a step of obtaining a buffy coat and a fraction with a heavier specific gravity than that of the buffy coat from the anticoagulated whole blood, and a step of promoting and activating an interaction between the obtained leukocytes and platelets at a given temperature for not less than a given time to cause secretion or release of a humoral factor from the leukocytes and/or platelets and performing a recoagulation treatment of blood components including the humoral factor with a re-coagulating agent.

The invention relates to a new method for in vitro expansion of CD4+CD25.sup.HighCD127.sup./LOWfoxP3+Tregs, wherein the process of Treg expansion takes place permanently or temporarily at a temperature below 37 C., optimally at a temperature of 33 C., the isolated Tregs are expanded in SCGM or X-vivo-20 medium supplemented with human serum or with foetal bovine serum, and magnetic beads coated with anti-CD3 and anti-CD28 antibodies at 1:1 (cell:bead) ratio and interleukin-2 are added to the culture.