The present invention provides for identification and use of small molecules to induce pluripotency in mammalian cells as well as other methods of inducing pluripotency.

The technology relates in part to methods and compositions for ex vivo proliferation and expansion of epithelial cells.

Formulations and methods to increase the production of recombinant proteins, and other aspects, are disclosed. The formulations and methods relate to increasing mannose or calcium concentration, or both, in a cell culture medium formulation for culturing cells that express recombinant proteins. In some embodiments, a mammalian cell culture medium formulation is provided that has at least one of mannose at about 3.5 g/L or more and calcium in a range from about 1.5 mM to about 9.5 mM. Numerous other aspects and/or embodiments are provided.

Disclosed are methods for treating or limiting development of diabetes, by transplanting into the eye of a subject with diabetes or at risk of diabetes an amount effective to treat or limit development of diabetes of insulin-producing cells engineered to reduce expression of a β3 subunit of Cav (Cavβ3).

A process of using a fish plasma component in a nutrient medium for cell culture includes obtaining a fish that is a progeny of domesticated broodstock that are reared under consistent and reproducible conditions. Blood is obtained from the fish, and plasma is separated from the blood. One or more specific components of the plasma are then extracted, and cells are cultured in a nutrient medium using the one or more extracted plasma components, and none of any remainder of the plasma. The plasma and/or the plasma components is/are tested for presence and/or level of endotoxin. Extracting the one or more specific components of the plasma, and/or culturing the cells is only performed if the testing indicates an endotoxin level below a predetermined threshold. The cells cultured using the extracted one or more plasma components are other than fish cells.

A method of producing a 3D tissue composite, comprising: a preparation step in which a multiple number of sheet-shaped first structures containing first cells are prepared, wherein at least one of the multiple number of first structures holds a second structure containing second cells; a stacking step in which the multiple number of first structures are stacked to form a 3D composite; and a culturing step in which the 3D composite is cultured to form a 3D tissue composite containing first tissues formed from the first cells and second tissues formed from the second cells.

Cells may be cultured by a method including the following steps: a first step of preparing a population of cell aggregates having a major axis of not more than 400 μm, and a second step of suspension culturing the population of cell aggregates obtained in the first step.

Herein the inventors demonstrate that mineralization is a natural ability of cells cultured with at least two elements: calcium and acyclic alkane phosphoester salt or inorganic phosphate salt. The present invention provides methods for producing hydroxyapatite (HAP) in cell culture by supplying cells with these elements. The natural HAP crystals produced by these methods may be utilized in biomedical applications such as bone grafting. Also provided are methods of measuring organic phosphates in a sample from a subject and methods of measuring the glycerophosphates in a sample from a subject.

Improved production of recombinant proteins in E. coli, reliant on tightly controlled autoinduction, triggered by phosphate depletion in stationary phase. The process also provides an optimized autoinduction media, enabling routine batch production at various culture volumes where cells densities routinely reach ˜5-7 g cell dry weight per liter and offer protein titers above 2 g/L. The methodology has been validated with a set of diverse heterologous proteins and is of general use for the facile optimization of routine protein expression from high throughput screens to fed-batch fermentation.

Methods and compositions for modifying glycans (e.g., glycans expressed on the surface of live cells or cell particles) are provided herein.