Delivering a payload across a plasma membrane of a cell includes providing a population of cells and contacting the population of cells with a volume of an aqueous solution. The aqueous solution includes the payload and alcohol content greater than 5 percent concentration. The volume of the aqueous solution may be a function of exposed surface area of the population of cells, or may be a function of a number of cells in the population of cells. Related compositions, apparatus, systems, techniques, and articles are also described.

The present invention discloses formulation of a serum-free medium used for human pluripotent stem cells, which comprises the following raw materials: inorganic salt components, organic components, amino acids and amino acid salts, energy substances and metabolic intermediates, vitamins and antioxidants, proteins and polypeptides, trace elements and chromogenic substances; while the culture process comprises the following steps: selecting a basic formulation, performing combination screening, identifying and evaluating results, and testing a new formulation of culture; and proportioning according to the following methods: adding aforesaid raw materials into 950 ml of water for injection, stirring gently until dissolved, and finally adding 2.438 g of sodium bicarbonate, and stirring gently until dissolved, and then adding 1 liter of water for injection, adjusting the pH to the desired value with 1 mol/L sodium hydroxide solution or 1 mol/L hydrochloric acid solution, finally filtering sterilized with 0.1 m diameter filter under positive pressure, and storing the medium solution in dark place at 2 C.-8 C., the invention solves the problem of high cost of domestic import of serum-free formulation.

A novel culture media formula that is thoroughly optimized to support high growth rate under low seeding density conditions, require minimal media exchanges, and at low cost, while maintaining differentiation reproducibility is provided. This formula is capable of supporting both human induced pluripotent stem cell (hiPSC) generation and culture for >100 passages. Generation of B8 supplement aliquots suitable for making 100 liters of media is simple for any research lab with basic equipment, with complete bottles of media costing $12 USD per liter. Weekend free hiPSC cell culture methods are possible with this formulation.

A method for growing polarized endometrial cells, said method comprising: (a) disposing endometrial cells on a scaffold, said scaffold comprising a silica-based glass composition, characterized by multi-modal porosity, said scaffold being to define a top side and a bottom side; (b) providing nutrients to said top and bottom sides of said scaffold and an environment to grow polarized endometrial cells on said scaffold.

Axenic media and methods for growing E. Chaffeensis and/or A. phagocytophilum are provided. In general, the axenic media includes intracellular phosphate buffer (IPB), a carbon source, FBS, a mixture of amino acids, and at least one further component selected from the group consisting of glucose 6-phosphate (G6P), ATP, DTT, GTP, UTP, CTP, and any combination thereof.

Methods and compositions are provided for generating targeted genetic modifications on the Y chromosome or a challenging target locus. Compositions include an in vitro culture comprising an XY pluripotent and/or totipotent animal cell (i.e., XY ES cells or XY iPS cells) having a modification that decreases the level and/or activity of an Sry protein; and, culturing these cells in a medium that promotes development of XY F0 fertile females. Such compositions find use in various methods for making a fertile female XY non-human mammal in an F0 generation.

The present invention is directed generally to dry cell culture media or feeds in pellet formats which can be reconstituted into liquid media for culturing cells in vitro. Each pellet composition may comprise the same or a different composition; for example, different vitamins, amino acids, buffers, trace salts, pH, iron chelators, etc. The invention also relates to methods of making dry cell culture media by altering ratios of different pellet compositions, or, methods of making modular dry cell culture media, or customizing media formulations for growing a cell type using pellets. The media pellets may be easier to handle either before reconstitution, during shipping and handling; and/or during reconstitution. Media pellets may be used in any container like bags including sterile, single use bags for preparing media formulations. The invention also relates to kits and culture systems using media pellets.

By establishing effective methods for shrimp 3D cell culture and passage, the present invention provides a technology of continuous shrimp cell culture intended for the establishment of immortalized shrimp cell lines. The present invention provides a preparation method of matrigel for 3D cell culture of shrimp by optimizing an additive proportion of matrigel. The present invention further provides a technology of separation and 3D cell culture of shrimp haemolymph cells, where shrimp haemolymph cells adhere to and grow on the surface of the matrigel in the form of a single round cell and a cell pellet/cellular spheroid, with survival and growth abilities being superior to 2D culture effects. The above technology is achieved by optimizing a formula of complete medium for shrimp cells, selecting the medium as an anticoagulant and a diluent for shrimp haemolymph cells, selecting a 3D culture method for surface-adhered growth in the matrigel.

In certain embodiments, the present invention provides a method of purifying a monomeric monoclonal antibody from a mixture which comprises the monomeric monoclonal antibody and one or more contaminants, comprising: a) subjecting the mixture to cation exchange chromatography (CEX) matrix, wherein the monomeric monoclonal antibody binds to the CEX matrix; b) contacting the CEX matrix with a wash solution at a pH which is between about 7 and about 7.8; c) eluting the monomeric monoclonal antibody from the CEX matrix into an elution solution, thereby purifying the monomeric monoclonal antibody.

The present disclosure is directed to a method for cultivating a Bordetella species, comprising: cultivating a Bordetella species under aerobic conditions in a liquid culture medium; and maintaining a pH of the liquid culture medium by using a strong acid, such as nitric acid, or using a first and second acid, wherein the first acid is an inorganic acid that dissociates essentially completely in water, such as nitric acid, hydrochloric acid or sulfuric acid, and wherein the second acid is an inorganic acid having an acid dissociation constant (pKa) of greater than 1, such as phosphoric acid. Methods for increasing the yield of Bordetella finbria agglutinogen 2 and fimbrial agglutinogen 3 (FIM2/3) in a supernatant fraction from a Bordetella culture are also provided.