Micro-Organosphers, including Patient-Derived Micro-Organospheres (PMOSs), apparatuses and methods of making them, and apparatuses and methods of using them. Also described herein are methods and systems for screening a patient using these Patient-Derived Micro-Organospheres, including personalized therapies.

A method for culturing primary gynecological tumor cells and culture medium used therein. The method includes using mild cell dissociation reagents to treat a gynecological solid tumor tissue, to ensure the vitality of tumor cells in the tissue to the greatest extent; preparing a special serum-free medium, and using a suspension culture system to culture solid tumor cells derived from a gynecological tumor in vitro to ensure the normal expansion of tumor cells and eliminate interference from normal cells to the greatest extent. The primary gynecological tumor cell culture obtained by the method of the present invention can be used for various cell-based in vitro experiments, second-generation sequencing, construction of animal models, construction of cell lines and the like. It is foreseeable that this culture method has broad application prospects in the fields of gynecological tumor research and clinical diagnosis and treatment.

A composite material film for expanding circulating tumor cells ex vivo and a preparation method thereof, a kit and a method for expanding circulating tumor cells ex vivo, a method for detecting an effect of a drug, and a cryopreservation solution are provided. The preparation method includes: mixing one or more kinds of particles and a solvent to form a mixed liquid, in which the particles are selected from the group consisting of metal particles, metal oxide particles, silicon oxide particles and combinations thereof; placing the mixed liquid on a substrate to form a particle layer; adding a medium material to the particle layer, in which the medium material is selected from the group consisting of styrene and its derivatives, polyester monomers, silicon oxide compounds and combinations thereof; and polymerizing the medium material to form a medium layer to fix the particle layer on the substrate.

A drug screening platform simulating hyperthermic intraperitoneal chemotherapy including a dielectrophoresis system, a microfluidic chip and a heating system is disclosed. The dielectrophoresis system is used to provide a dielectrophoresis force. The microfluidic chip includes a cell culture array and observation module and a drug mixing module. The cell culture array and observation module are used to arrange the cells into a three-dimensional structure through the dielectrophoresis force to construct a three-dimensional tumor microenvironment. The drug mixing module is coupled to the cell culture array and observation module and used to automatically split and mix the inputted drugs and output the drug combinations into the cell culture array and observation module. The heating system is used for real-time temperature sensing and heating control of the drug combinations on the microfluidic chip to simulate high-temperature drug environment when performing hyperthermic intraperitoneal chemotherapy on the three-dimensional tumor microenvironment.

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 are compositions and methods for production of anti-inflammatory cytokines, growth factors, or chemokines. Provided are nucleic acids (e.g., expression vectors) that include an NFκB inflammation response element operably linked to a nucleotide sequence encoding an anti-inflammatory cytokine (e.g., IL-4). In some cases, the nucleic acid is an expression vector selected from: a linear expression vector, a circular expression vector, a plasmid, and a viral expression vector. Also provided are cells (e.g., mesenchymal stem cells—MSCs) comprising a nucleic acid that includes an NFκB inflammation response element operably linked to a nucleotide sequence encoding an anti-inflammatory cytokine. In some cases, the nucleic acid is integrated into the cell's genome. Also provided are methods for treating an individual having an inflammation-associated ailment, which can include administering an MSC to the individual, where the MSC includes an NFκB inflammation response element operably linked to a nucleotide sequence encoding an anti-inflammatory cytokine.

In certain example embodiments, the invention provides a method of generating an ex vivo cell-based system comprising dissociating an original tissue sample obtained from a subject into a single cell population; determining an in vivo phenotype of the tissue sample by conducting single-cell RNA analysis on a first portion of the single cells; establishing an ex vivo cell-based system from a second portion of the single cells; and culturing the ex vivo cell-based system in a medium or conditions selected to maintain the in vivo phenotype. In some embodiments, the original tissue sample is a tumor tissue sample, such as a pancreatic ductal adenocarcinoma (PDAC) tumor sample.

The present invention provides a pharmaceutical composition for the treatment of patients having ovarian cancer, lung cancer or mesothelioma and showing a Selection Factor of −30% or below, comprising a therapeutically effective amount of ipilimumab, and optionally a pharmaceutically acceptable diluent or carrier, wherein the patient is selected on the basis of a positive response to an ex vivo three-dimensional (3D) patient derived tumour culture, the method comprising: (a) preparing a three-dimensional, optionally size-normalised, tumour culture from a patient-derived tumour sample in a multitude of replicates; (b) adding one or more immunotherapeutic agents to the culture, and (c) culturing for a pre-defined time period; and (d) determining the effect that the one or more immunotherapeutic agents has on the tumour cell aggregates by measuring the total area of objects in the culture that are above a threshold area associated with tumour cell aggregates, and the total area of objects that are below a threshold associated with immune cells, using three-dimensional imaging of the cell culture; wherein if following culturing with a composition comprising ipilimumab, the total area of the large objects decreases and/or the total area of the small objects increases relative to a control the patient is treated with ipilimumab.

Engineered tissue models based on three-dimensional (3D) scaffolds, also referred to herein as tissue-engineered 3D models, can be used as in vitro diagnostic and drug screening tools for predicting, preventing and/or treating cancer metastases.

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.