Disclosed herein include methods and compositions for culture medias for in vitro culture of synthetic embryos from mammalian pluripotent stem cells and extra-embryonic stem cells. The methods and compositions described herein can generate synthetic embryos at different developmental stage reaching early organogenesis and beyond. Disclosed herein also include an embryo culturing system and methods of using same.

Disclosed herein are insulin resistance reporters for use in quantifying insulin response in biological cells. These biological cells may be stem cell compositions or derivatives thereof comprising the insulin resistance reporter. The stem cell derivatives include but are not limited to insulin responsive cells, tissues, or organoids, such as pancreatic, brain, adipose, muscle, or liver cells, or tissues or organoids thereof. Also disclosed herein are methods of using said insulin resistance reporters and cells with these insulin resistance reporters as models to examine insulin resistance and screening for compounds that are potentially useful for the treatment of diseases or disorders associated with insulin resistance. The cells comprising an insulin resistance reporter may be hepatic cells or liver organoid compositions, which can be used in investigating hepatic insulin resistance, for example, as a result of non-alcoholic fatty liver disease or steatohepatitis.

The present disclosure describes a microfluidic chip for culturing and in vitro testing of 3D organotypic cultures. The tests may be performed directly on the organotypic culture in the microfluidic chip. The microfluidic chip includes at least one microfluidic unit which includes two fluidic compartments, such as upper and lower, separated by a permeable supporting structure, one or more access opening for the fluidic compartments, and a set of lids interchangeable with a set of insets. The permeable support structure serves as a support for the organotypic culture. The upper and lower compartments may include inlets and outlets which allow fluids to be perfused into the lower compartment and fluids to be perfused into the upper compartment. The access opening may be closed with a lid or accommodate an inset.

Disclosed are a spheroid array and more particularly, a droplet trapping structure array capable of isolating all or selected spheroids into an isolated droplet array environment and the use thereof. The droplet-trapping structure array and the method and device for transferring spheroids using the same have the advantages of transferring droplets or spheroids with very high efficiency and very small variation between users by simply contacting two arrays. The spheroid transfer method and device enable mass-production of spheroid arrays in an isolated environment. In particular, the droplet trapping structure array and the spheroid transfer method can be useful for the treatment of spheroids with various reagents and the exchange of culture media.

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

The present invention relates to a method, which patterns 3D organoids prepared from human pluripotent stem cells and chops the same so as to culture oligodendrocyte progenitor cells, and induces the differentiation thereof so as to obtain a large quantity of finally differentiated oligodendrocytes. Compared to cells differentiated by a conventional differentiation method, oligodendrocytes obtained in a large quantity have the same or superior reproducibility, stability, and functionality and have remarkably shortened differentiation time, and thus are expected to be very useful for cell therapeutic agents or for screening for therapeutic drugs.

The present disclosure provides a method of bioprinting a 3-D structure comprising one or more biologically-relevant materials on a super-hydrophobic surface. In one embodiment, the method comprises providing a composition having one or more biologically-relevant materials dispersed within a biocompatible medium. A pattern comprising a hydrophilic material is deposited on a defined area of the super-hydrophobic surface, wherein the pattern is modeled after a biological structure. The composition having the one or more biologically-relevant materials is then bioprinted atop the hydrophilic surface to form a 3-D structure, wherein the hydrophilic surface maintains the 3-D structure in a desired position or shape on the super-hydrophobic surface.

A method of obtaining a pluripotent-like multipotent cell, including providing a cell of a first type which is not a pluripotent-like multipotent cell; contacting the cell of a first type with an agent capable of remodeling the chromatin and/or DNA of the cell; transiently increasing expression of at least one pluripotent gene regulator in the cell of a first type, to a level at which the at least one pluripotent gene regulator is capable of driving transformation of the cell of a first type into the pluripotent-like multipotent cell; and placing or maintaining the cell in a differentiation medium and maintaining intracellular levels of the at least one pluripotent gene regulator for a sufficient period of time to allow a stable pluripotent-like multipotent cell to be obtained; wherein the pluripotent-like multipotent cell so obtained does not exhibit teratoma formation in vivo.

The present invention discloses, in one embodiment, a method of using human induced pluripotent stem cells to generate three-dimensional human organ tissue for therapeutic drug toxicity and discovery⋅. In one embodiment, a high throughput microtiter plate is loaded with both wild type and Rett disease 3D spheroids and exposed to a drug library, and activity is measured and analyzed for disease rescue to wild type cell behavior.