The present disclosure provides organoid co-cultures and methods of using such co-cultures. In particular, the present disclosure provides organoid-immune cell and organoid-bacterial cell co-cultures. The present disclosure further provides methods for testing therapeutic agents using the disclosed organoid co-cultures.

Described herein are methods for providing an in vitro intestinal model system, e.g., using primary cells instead of cell lines and/or cancerous cells.

A primary cell culture medium for culturing primary lung cancer epithelial cells, a culture method using the primary cell culture medium, and an application thereof in drug efficacy evaluation and screening. The culture medium contains an MST1/2 kinase inhibitor, a ROCK kinase inhibitor, a fibroblast growth factor, at least one additive selected from a B27 additive and an N2 additive, an epidermal growth factor, transferrin, gastrin, and a TGF type I receptor inhibitor.

The present invention relates to a method for constructing a human pluripotent stem cell-derived liver organoid having enhanced drug metabolic potentials and a liver organoid constructed by the method. Because differentiation is made from human induced pluripotent stem cells dedifferentiated from somatic cells of patients, the cells are easily supplied so that patient-specific liver organoids can be prepared. The organoids can similarly simulate the real human liver structure because they consist of hepatocytes, cholangiocytes, gallbladder cells, and microduct structures, have excellent expression and functions of drug metabolism enzymes, and exhibit drug toxicity, drug metabolic capability, and drug-induced cardiotoxicity. Thus, the organoids can be advantageously used as liver models for searching for pathogenesis of liver diseases and for evaluating drug stability.

The invention relates to a method for culturing epithelial stem cells, isolated tissue fragments comprising the epithelial stem cells, or adenoma cells, and culturing the cells or fragments in the presence of a Bone Morphogenetic Protein (BMP) inhibitor, a mitogenic growth factor, and a Wnt agonist when culturing epithelial stem cells and isolated tissue fragments. The invention further relates to a cell culture medium comprising a BMP inhibitor, a mitogenic growth factor, and a Wnt agonist, to the use of the culture medium, and to crypt-villus organoids, gastric organoids, pancreatic organoids, liver organoids, colon organoids, Barrett's Esophagus organoids, adenocarcinoma organoids and colon carcinoma organoids that are formed in the culture medium.

The invention provides for methods of differentiating pancreatic endocrine cells into pancreatic beta cells expressing PDX1, NKX6.1, MAFA, UCN3 and SLC2A. These pancreatic beta cells may be obtained by step-wise differentiation of pluripotent stem cells. The pancreatic beta cells exhibit glucose-dependent mitochondrial respiration and glucose-stimulated insulin secretion similar to islet cells.

A culture medium for primary ovarian cancer cells, an in vitro culture method and an application thereof. The culture medium contains: an MST1/2 kinase inhibitor; a Rho kinase inhibitor, which is selected from at least one among Y27632, Fasudil and H-1152; an insulin-transferrin-selenium supplement; insulin; prostaglandin E2; an epidermal growth factor; gastrin; an insulin-like growth factor-1; cholera toxin; amphiregulin; N2; and B27. Compared with existing culture methods, the in vitro culture using said culture medium has higher amplification efficiency. Using the culture medium for the culturing of primary ovarian cancer cells may maintain the morphological structure and pathological features of primary tissues, and improve the success rate and survival rate of the cultured primary ovarian cancer cells.

The invention relates to a method for culturing epithelial stem cells, isolated tissue fragments comprising the epithelial stem cells, or adenoma cells, and culturing the cells or fragments in the presence of a Bone Morphogenetic Protein (BMP) inhibitor, a mitogenic growth factor, and a Wnt agonist when culturing epithelial stem cells and isolated tissue fragments. The invention further relates to a cell culture medium comprising a BMP inhibitor, a mitogenic growth factor, and a Wnt agonist, to the use of the culture medium, and to crypt-villus organoids, gastric organoids, pancreatic organoids, liver organoids, colon organoids, Barrett's Esophagus organoids, adenocarcinoma organoids and colon carcinoma organoids that are formed in the culture medium.

The invention relates to improved culture methods for expanding epithelial stem cells and obtaining organoids, to culture media involved in said methods, and to uses of said organoids.

Described herein are methods for providing an in vitro intestinal model system, e.g., using primary cells instead of cell lines and/or cancerous cells.