A method of producing induced exosomes, the method comprising: contacting an isolated population of stem cells with an amount of a prostaglandin E receptor 4 (EP4) antagonist effective for inducing release of exosomes, whereby induced exosomes are released from the stem cells, and isolating the induced exosomes.

Disclosed are methods of producing organoids in the absence of any exogenous extracellular matrix or in the presence of an exogenous extracellular matrix at a concentration lower than gelling concentration, using microcontainers sealed with a hydrogel lid as well as organoids produced by this technique.

The invention provides tissue culture system for primary cells (e.g. normal mammalian primary epithelial progenitors). This system includes: a) a serum-free, chemically defined cell culture media; and, b) methods for isolation and in vitro long-term propagation of primary cells (e.g. primary epithelial cells). Primary cells so isolated and cultured can be kept undifferentiated and proliferate for many weeks (>15 weeks) or population doubling (>35 PD) without senescence, or any detectable genetic alterations. Upon changing media/culture conditions, these cells can be induced to differentiate. The invention also provides methods to transform normal primary cells so cultured into “cancer stem cells.” The genetically defined cancer stem cell tumor model mimics the behavior of the disease closely, e.g., the cells are invasive, hormone responsive and metastatic when injected into mice. The tumor cells express genes that are specific to cancer stem cells identified in patient samples.

Provided is a culture medium containing amphiregulin for culturing primary breast epithelial cells and a culture method involving using the medium. In the culturing method, primary cells are cultured on a culture vessel coated with an extracellular matrix gel by using the culture medium, and the primary cells grow on the culture vessel, which has been coated with the extracellular matrix gel, and proliferate rapidly under the combined action of nutrient factors and an extracellular matrix contained in the primary cell culture medium. The cell model obtained by the primary cell culture medium and the primary cell culture method can be used for the evaluating the efficacy of and screening drugs.

The invention relates to methods for generating multipotent mammary stem cells from isolated and cultured human breast luminal cells. The method comprises the steps: 1. isolating and growing normal differentiated cells in vitro; 2. treating the differentiated cells with either a conditioned medium from active fibroblasts or cytokine. The invention also relates to multipotent mammary stem cells, cultures of the multipotent stem cells, differentiated cells, tissues, organs derived from the culture multipotent stem cells isolated by the methods disclosed and therapeutic and other uses for those cells thereof.

This invention relates to live cell constructs for producing milk in culture and compositions comprising a milk product produced by the live cell contracts, as well as methods for making a live cell construct for producing milk in culture, methods of producing milk in culture, and methods of producing a modified primary mammary epithelial cell or an immortalized mammary epithelial cell for use in a live cell construct and other methods of the present invention.

The disclosure relates to methods, systems and compositions for use in the production of milk. More specifically, the disclosure is directed to systems, compositions and methods for in-vitro production of milk using an array of mammary organoids seeded on tertiary-branched, resilient duct scaffolding.

The present disclosure provides a method for inducing transdifferentiation of somatic cells into mammary epithelial cells in vitro using a small molecule compound. The method includes: inhibiting an expression of TGFbeta R1 and related sites thereof, to induce the transdifferentiation of the somatic cells into the mammary epithelial cells in vitro. The present disclosure fills a gap in the technology of inducing the transdifferentiation of fibroblasts to the mammary epithelial cells using the small molecule compound; the present disclosure also provides a research platform for in vitro researches on a mammary gland bioreactor, mammary gland development and differentiation, breast cancer, and transdifferentiation of the fibroblasts into other types of functional cells.

A method for producing a mammalian milk like product, for example a human milk like product comprising generating lactocytes derived from mammalian induced pluripotent stem cells (miPSC), for example human induced pluripotent stem cells (hiPSC), and expressing the mammalian milk like product, for example the human milk like product from lactocytes.

The present invention relates to the fields of life sciences and cell and tissue cultures, especially 3D cultures. Specifically, the invention relates to a method of maintaining the presence or activity of a human or mouse estrogen receptor (ER) in a cell of an ex vivo mammary cell or tissue culture or in a cell of other hormone responsive cell or tissue culture. Also, the present invention relates to a method of maintaining a luminal epithelial phenotype and/or cell identity of a mammalian cell in an ex vivo cell or tissue culture. Still, the present invention relates to a 3D matrix or 3D medium comprising the matrix for ex vivo culture, wherein said 3D matrix or 3D medium comprises one or more mammalian cells or tissues embedded in said 3D matrix or 3D medium, and to a system for ex vivo culture, wherein the system comprises mammalian cells or tissues embedded in a 3D matrix or 3D medium comprising said matrix. Still furthermore, the present invention relates to use of the 3D matrix, 3D medium or system of the present invention e.g. for ex vivo culture of a mammalian cell, drug discovery methods, biomarker studies and/or estrogen receptor (ER) signaling studies.