The present disclosure provides a method of producing uniformly sized organoids/multicellular spheroids using a microfluidic device having an array of microwells. The method involves several successive steps. First, a microfluidic device containing parallel rows of microwells that are connected with a supplying channel is filled with a wetting agent. The wetting agent is a liquid that is immiscible in water. For example, the wetting agent may be an organic liquid such as oil. In the next step, the agent in the supplying channel and the microwells is replaced with a suspension of cells in an aqueous solution that contains a precursor for a hydrogel. Next, the aqueous phase in the supplying channel is replaced with the agent, which leads to the formation of an array of droplets of cell suspension in the hydrogel precursor solution, which were compartmentalized in the wells. The droplets are then transformed into cell-laden hydrogels. Subsequently, the agent in the supplying channel is replaced with the cell culture medium continuously flowing through the microfluidic device and the cells within the hydrogels are transformed into multicellular spheroids.

The present invention provides improved methods and media for culturing stem cells and propagating organoids. In particular, the methods provide improved methods for culturing normal and tumour or cancer stem cells and their propagation into organoids. Specifically provided are suspension methods with particular utility for scalable organoid expansion, biobanking, experimental manipulation, miniaturization and imaging based high-throughput drug screening.

The present disclosure relates to a method for controlling freeze-drying of a hydrogel wherein the method comprises adjusting the residual water content of the freeze-dried hydrogel, and to a freeze-dried hydrogel composition comprising nanoscale cellulose and a at least one biomolecule selected from an oligosaccharide or a disaccharide. The present disclosure further concerns the use of the freeze-dried hydrogel composition for cell culturing, a cell culturing scaffold as well as a process for manufacturing a reconstituted hydrogel.

The present disclosure provides a substrate for cell culture. Systems comprising the substrate, and methods for using and manufacturing the substrate are also disclosed herein.

The present disclosure provides for a cell stabilizing medium which comprises gelatin. The cell stabilizing medium help maintain cell viability, e.g., after thawing of a biological material post-cryopreservation.

The present specification provides methods for augmenting the antigenic content, especially of tumor-associated antigens (TAA), and immunogenicity of cancer cells; methods for enhancing cross-presentation in dendritic cells, compositions comprising such manipulated cells derived from single cancer patients; and methods of using those compositions as a personal immunotherapeutic product to treat the donor patient's cancer.

Provided are a culture medium and cultivation method of rapidly expanding primary cells of esophageal squamous carcinoma in vitro, and the use thereof in screening drugs. The culture medium comprises an initial culture medium selected from DMEM/F12, DMEM, F12, or RPMI-1640, a Rho protease inhibitor, an antibiotic, insulin, an N2 additive, insulin-like growth factor 1, a non-essential amino acid, and optionally, hydrocortisone, optionally, glutamine, and optionally, bovine pituitary extract.

The present invention relates to thin filters comprising melt electro spinning writing (MEW) fibers for capturing and culturing circulating tumor cells (CTCs). The invention further relates to processes for producing the filters, methods for capturing and culturing CTCs using the filters, kits and devices comprising the filters and uses of the filters.

The subject matter of the present invention is a method for differentiating epithelial stem cells, comprising culturing one or more epithelial stem cells in contact with an extracellular matrix in the presence of an expansion medium, a bovine pituitary extract, a receptor tyrosine kinase ligand, a supernatant of primary fibroblasts and optionally, a Rho kinase inhibitor.

The present disclosure provides methods of evaluating a therapeutic agent for cancer, and methods of cancer treatment.