This disclosure generally relates to cell-based therapies for treatment of visual disorders, including disorders of the cornea. Methods are exemplified for directed differentiation of corneal cells from stem cells. Compositions of corneal endothelial cells and uses thereof are also provided. Exemplary compositions exhibit improved cell density and/or more youthful gene expression relative to cells obtained from donated tissue.

Methods and compositions are described for making phenotypically female fertile animals from XY donor cells and suitable host embryos. Culture media and methods are provided for maintaining XY donor cells in culture that after introduction into a host embryo and gestation in a suitable host will result in fertile XY female animals. Methods and compositions are described for making fertile female animals in an F0 generation from a donor XY cell and a host embryo, as are methods for making F1 progeny that are homozygous for a modification from a heterozygous F0 fertile male and a heterozygous F0 fertile female sibling.

The present disclosure provides methods of generating germ layers from stem cells comprising culturing the stem cells in a culture media having osmolality ranges that promote the generation of specific germ layer progenitor cells. The present disclosure also includes a method to generate different cell lineages from the germ layers as well as to detect them by immunological methods. The present disclosure further provides methods for the generation, isolation, cultivation and propagation of committed progenitor cells and for the production of differentiated cells from the three germ layers. The present disclosure also provides culture media and kits for use in inducing the three germ layers.

The present invention relates to the fields of medical microbiology and vaccines. In particular the invention relates to a process for detergent-free preparation of outer membrane vesicles (OMV) of Gram negative bacteria for use in vaccines, to OMV obtainable by said process, and to a pharmaceutical composition comprising such OMV. The present invention further relates to the use of OMV of the present invention as a medicament in particular for use in a method for eliciting an immune response.

The present invention provides a compounded cell culture medium powder formulation comprising: a basal medium powder and a cell culture media supplement, wherein the cell culture media supplement comprises and one or more salts; one or more growth factors; one or more inorganic ions; an amino acid supplement comprising one or more of asparagine, glutamine, histidine, and serine; one or more buffers; and one or more anti-foaming agents. The invention further provides methods of making a compounded cell culture medium powder formulation methods of making a cell culture medium for growing mammalian cells and methods of producing a protein of interest by culturing cells in the cell culture medium and isolating the protein of interest.

The present invention pertains to methods of producing polypeptide of interest in cell cultures lacking glutamine. The present invention further pertains to a method of producing a protein of interest in a large scale cell culture, comprising supplementing the cell culture with nucleic acid synthesis precursors and/or corticosteroids.

The invention relates to methods of differentiation, isolation, propagation, and storage of small colony variants (SCVs) of E. coli, preferably E. coli 83972 or E. coli HU2117, or modified or variant forms thereof, and methods for using the pre-pared SCV bacteria to establish probiotic biofilms in treated subjects and/or on treated medical devices.

Disclosed are a method for preparing a target recombinant protein with modulated galactosylation or a method for modulating the galactosylation of a target recombinant protein, including increasing the osmolality of a culture solution of animal cells which express a target recombinant protein during the animal cell culture; a method for preparing a target recombinant protein with modulated galactosylation or a method for modulating the galactosylation of a target recombinant protein, including supplementing asparagine to a culture solution of animal cells which express a target recombinant protein during the animal cell culture; a method for preparing target recombinant protein with modulated galactosylation or a method for modulating the galactosylation of a target recombinant protein, including increasing the osmolality of an animal cell culture which expresses a target recombinant protein and supplementing asparagine thereto during the animal cell culture; and a target recombinant protein with modulated galactosylation, which is prepared by the method.

Disclosed is a hydrogel composition loaded with a base that regulates and maintains the pH of cell culture media. The invention particularly discloses in situ pH managing hydrogels (pHmH) loaded with base Mg(OH).sub.2 for pH control in small scale cultures comprising adherent mammalian cells, suspended mammalian cells, microorganisms, a microorganism with plasmids and the like thereof.

An apparatus and method for the delivery and deposition of particles air-liquid-interface (ALI) cell cultures includes a sample inlet coupled to a growth tube having interior walls that are wet. The growth tube is configured to operate at a first temperature along a first length of the tube and a second temperature along a second length positioned between the first length and a growth tube outlet. The apparatus also includes a nozzle plate having a plurality of nozzles. The exposure chamber adapted to hold cell cultures at an air-liquid interface positioned underneath the plurality of nozzles and a temperature regulator adapted to control a temperature of the exposure chamber. The apparatus also includes a controller including instructions operable to cause the controller to maintain a relative humidity within the exposure chamber by controlling at least the second temperature of the growth tube and the temperature of the exposure chamber.