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.

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 provides methods of identifying and distinguishing different types of nerve cells in ex vivo cell culture, the method comprising the steps of: a) culturing somato-sensory nerve cells ex vivo, b) loading the nerve cells with a calcium, sodium, or voltage-sensitive indicator or expressing a genetically encoded calcium, sodium, or voltage-sensitive indicator, c) pulsing the nerve cells with an electrical train of bipolar square waves at two different voltages and two different frequencies; wherein the first voltage is 10 V/cm or less (low voltage) and the second voltage is between 12 and 20 V/cm (high voltage); and wherein the first frequency is 5 Hz or less (low frequency) and the second frequency is between 15 and 20 Hz (high frequency), and d) detecting activation of the nerve cell by measuring the changes in the signal intensity of the indicator, wherein low voltage and low frequency activation indicates a first type of cell and activation detected only at high voltage indicates a second type of cell.

The disclosure herein relates to construction and application of engineering bacteria capable of secreting and expressing diacetylchitobiose deacetylase, and belongs to the technical field of fermentation engineering. Firstly, recombinant B. subtilis capable of heterologously secreting and expressing a diacetylchitobiose deacetylase gene is constructed, and a signal peptide fragment yncM is added into the recombinant vector for the first time. The signal peptide can secrete the target protein diacetylchitobiose deacetylase outside the cells of the recombinant B. subtilis, and a mutant of the 5-end untranslated region is acquired, thereby significantly increasing the expression level of the target protein, and greatly simplifying the subsequent enzyme separation and purification steps. When the acquired diacetylchitobiose deacetylase is fermented and cultured in a fermentation medium for 50-60 h, the enzyme activity reaches a maximum of 1,548.7 U/mL, and the maximum yield of the diacetylchitobiose deacetylase is about 620 mg/L. Simultaneously, the method has the advantages of low production cost, mild production conditions, simple purification process steps, safe production operation and the like.

The present invention disclosed a method for preparing of collagen having regeneration and repair effects from Wharton's Jelly mesenchymal stein cells, comprising steps of culturing the Wharton's Jelly mesenchymal stein cells in a first medium for 16 to 24 hours; replacing the first medium with a second medium for culturing the Wharton's Jelly mesenchymal stein cells for 36 to 48 hours; collecting the Wharton's Jelly mesenchymal stein cells and adding a cell lysis solution to lyse the Wharton's Jelly mesenchymal stein cells for 0.5 to 2 hours, adding an inorganic salt solution to the cell lysis solution to obtain a mixing solution for further incubation at 4 C. for 24 to 48 hours; centrifuging the mixing solution and collecting a sediment, dissolving the sediment by a preservation solution to obtain a collage.

The present disclosure provides methods and compositions of matter directed to removing heavy metals, such as lead, from aqueous solutions by bioremediation. The methods use bacteria, which thrive in the presence of heavy metals to precipitate the heavy metals from the aqueous solution. In some embodiments, the bacteria comprise Bacillus licheniformis.

Provided herein are methods of improving rAAV production in mammalian cells via the manipulation of a concentration of potassium chloride present in the media in which the mammalian cells are cultured and/or in which the rAAV particles are produced. The concentration of potassium chloride present in such media may be manipulated by, for example, supplementing a base medium with additional potassium chloride either before, at the same time as, or after the cells are contacted (e.g., infected) with one or more viral vectors (e.g., recombinant Herpes Simplex Virus (rHSV) vectors). Some embodiments of the present invention further contemplate supplementing the base medium with additional potassium chloride and sodium chloride in combination.

A microbial inoculant for high-speed humification of organic waste and a preparation method thereof are provided. According to the present disclosure, the microbial inoculant is prepared from Geobacillus stearothermophilus GT1, Thermus tengchongensis GT2, and Thermus amyloliquefaciens GT3. According to the present disclosure, the microbial inoculant prepared from hyperthermophiles with different functions directionally degrades proteins, promotes humification, and degrades complex polysaccharides and organic waste. The resulting products contain rich surface oxygen-containing functional groups, which can adsorb heavy metals and reduce the heavy metal migration rate. Meanwhile, the preparation method of the microbial inoculant provided by the present disclosure can solve the problem that multiple microbial strains are difficult to enrich, and materials for preparing the microbial inoculant can be recycled, which are environmentally friendly and free of secondary pollution.

Described herein are methods for enhancing the nuclear reprogramming of somatic cells to become induced pluripotent stern cells. In particular, the methods disclosed herein involve the use of damage-associated molecular pattern molecules (DAMP). In certain embodiments the DAMPs are aluminum compositions such as aluminum hydroxide. Such DAMPs have unexpectedly and surprisingly been found to enhance the nuclear reprogramming efficiency of the reprogramming factors commonly used to induce somatic cells to become induced pluripotent stern cells. Accordingly, this disclosure describes methods of nuclear reprogramming as well as cells obtained from such methods along with therapeutic methods for using such cells for the treatment of disease amendable to treatment by stem cell therapy; as well as kits for such uses.

Provided herein, inter alia, are compositions and methods for culturing mammalian cells. In certain aspects, the composition is a medium containing one or more of a lithium ion source, one or more fatty acids, and/or ethanol. Use of any of the cell culture media described herein to culture cells that have been genetically engineered to produce one or more recombinant polypeptides (for example, antibodies) can result in increased titers, a more favorable glycosylation profile, and/or modulated (e.g. decreased) amounts of high and low molecular weight species, and/or modulated (e.g. decreased) amounts of acidic or basic charge variants, compared to cells cultured in a medium that does not contain one or more of a lithium ion source, one or more fatty acids, and/or ethanol.