The disclosure pertains to novel eukaryotic cell suitable for recombinant production of a product of interest, wherein the effect of the expression product of an endogenous gene C12orf35 is impaired in said cell, thereby increasing their productivity when recombinantly expressing a polypeptide of interest. Furthermore, the present disclosure provides associated technologies wherein such host cells are used in recombinant production technologies.

The present invention provides a recombinant eukaryotic cell expressing one or more heterologous double strand break (DSB) repair proteins in an amount effective for enhancing DSB repair in the cell. The recombinant eukaryotic cell may express a recombinant product of interest. Also provided are methods for enhancing double strand break (DSB) repair in eukaryotic cells, establishing host cells for production of a recombinant product of interest, producing a recombinant product of interest, improving production of a recombinant product of interest by eukaryotic cells, and/or investigating suitability of eukaryotic cells as host cells for producing a recombinant product of interest.

The present invention relates to a method for reducing the oxidation of methionine residues in a recombinant protein by culturing cells expressing the recombinant protein in a cell culture medium and feeding the cells with glutamine.

The present invention is directed generally to systems and methods suitable for high level expression of recombinant proteins in suspension CHO cells. In particular, the invention allows introduction of the invention obviates the need to replace, replenish or supplement the growth medium during the procedure. The invention also relates to compositions and kits useful for culturing and transforming/transfecting suspension CHO cells.

Provided herein are methods of producing a recombinant protein that include fed-batch culturing a mammalian cell.

Recombinant expression vectors are disclosed that include a control sequence for recombinant expression of proteins of interest; the control sequence combines a mCMV enhancer sequence with a rat EF-1alpha intron sequence. Some of the vectors are useful for tetracycline-inducible expression. Some of the vectors contain a 5′ PiggyBac ITR and a 3′ PiggyBac ITR to promote genomic integration into a host cell chromosome. A method of selecting a stable production cell line for manufacturing a protein of interest is also disclosed. Also disclosed are mammalian host cells comprising the inventive recombinant expression vectors and a method of producing a protein of interest, in vitro, involving the mammalian host cell.

Provided herein are methods to increase the culture density and/or thickness of a cellular biomass in a cultivation infrastructure, to improve the culture of cells in the absence of serum in a cultivation infrastructure, and to promote anchorage-independent growth of a cellular biomass in a cultivation infrastructure. The methods comprise inhibiting the HIPPO signaling pathway, for example, by activating YAP1, activating TAZ, and/or inhibiting MOB1, LATS1 kinase, LATS2 kinase, WW45, MST1 kinase, and/or MST2 kinase in the cellular biomass. In some embodiments, the cellular biomass is harvested from the cultivation infrastructure for the formulation of cell-based food products or ingredients, such as animal meat manufactured from cells in an ex vivo process or for therapeutic applications such as organ or tissue transplantation or grafting.

A three-dimensional culture method for large-scale preparation of stem cells, comprising a three-dimensional microcarrier-based cell resuscitation method, a three-dimensional microcarrier cell culture-based in situ passage method, a three-dimensional microcarrier in situ freeze preservation method for cells, a three-dimensional microcarrier cell adsorption culture method, a method for harvesting cells on a three-dimensional microcarrier, a method for sampling cells cultured on a microcarrier, and a three-dimensional microcarrier-based cell large-scale expansion method.

A production method for pluripotent stem cells is provided, the method including the following (a) and (b): (a) a process filling a culture container with a liquid medium and thereafter raising, the temperature of the liquid medium in the culture container to the temperature at which pluripotent stem cells can proliferate; and (b) a process seeding pluripotent stem cells in the liquid medium in the culture container and culturing the pluripotent stem cells in suspension.

Methods of and system for growing and maintaining an optimized/ideal active yeast solution in the yeast tank and fermenter tank during the fermentation filling cycle are provided. A new yeast stage tank is used between the yeast tank and the fermenter tank allowing yeast to rapidly produce a huge amount of active young yeast cells for a fermenter during the filling period. A measurable and useful controlling factor, % DT/% Yeast by weight ratio (or “food” to yeast ratio), is used (e.g., % DT=glucose), which offers information on the health status of the yeast. The controlling factor is used to control the status of the yeast throughout the entire process.