A method for expanding a population of γδ T-cells is provided in which isolated activated Peripheral Blood Mononuclear Cells (PBMCs) are cultured in a medium comprising transforming growth factor beta (TGF-β) under conditions in which the production of effector γδ T-cells having therapeutic activity against malignant disease is favored. The use of TGF-β in the production of effector cells in particular Vγ9Vδ2 T-cells is also described and claimed.

The present invention relates to processes to make neosaxitoxin, and analogues and variants thereof, and intermediates in the production of neosaxitoxin in recombinant host cells. Neosaxitoxin and the analogues and variants thereof may be used in the production of pharmaceutical compositions.

The present disclosure provides a pharmaceutical composition for treating Alzheimer’s disease, which at least includes an extracellular vesicle that is prepared by a method including: a first culturing step: performing an amplification culture of an adipose-derived stem cell in a first culture medium at a cell density of 6,000-15,000 cells/cm.sup.2 until an amplification amount of the adipose-derived stem cell is above 90% of that before the culture; a second culturing step: culturing the amplified adipose-derived stem cell in a second culture medium at a cell density of 10,000-100,000 cells/cm.sup.2 for 20-30 hours; and an extracellular vesicle separating step: collecting a culture solution and separating the extracellular vesicle from the culture solution by utilizing a tangential flow filtration (TFF) or ultrafiltration method.

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.

The present invention relates to a method for promoting growth of gas-fermented microorganisms, comprising the following steps: S1. ultrasonically blending a surfactant and a culture medium, then adding a fluorine-containing alkyl compound to a mixture, and ultrasonically processing the mixture to obtain a perfluorocarbon nanoemulsion; S2. inoculating a bacterial suspension into the perfluorocarbon nanoemulsion, and introducing simultaneously a mixed gas to obtain a precursor; S3. cultivating the precursor in a shaker, wherein the precursor has a diameter of bubbles of 2.0-4.2 mm, and the bubbles have the total volume of less than 40 ml. Under these conditions, the microorganisms have a high utilization rate of the gas, can grow and metabolize more quickly, and can obtain bacteria and products more quickly. Compared with a traditional culture system, addition of a mass transfer material can promote gas mass transfer. Therefore, the gas consumption is small, and the cost is low.

A method for expanding a population of γδ T-cells is provided in which isolated activated Peripheral Blood Mononuclear Cells (PBMCs) are cultured in a medium comprising transforming growth factor beta (TGF-β) under conditions in which the production of effector γδ T-cells having therapeutic activity against malignant disease is favored. The use of TGF-β in the production of effector cells in particular Vγ9Vδ2 T-cells is also described and claimed.

According to the present disclosure, there is provided a method for producing stem cells including: preparing somatic cells; preparing a chimeric virus including a virus-derived genomic RNA harboring an inducer RNA that induces somatic cells into stem cells and a virus-derived envelope surrounding the genomic RNA, wherein the genomic RNA and the envelope are derived from different viruses; and introducing the inducer RNA into the somatic cells using the chimeric virus.

A method, a medium and a kit for the enrichment and detection of Listeria species, especially Listeria monocytogenes. The medium is an enrichment medium of C12 to C16 fatty acids and/or derivatives thereof.

Methods for maintaining and expanding human CD34+ hematopoietic stem cells (HSCs) are provided using chemically-defined culture media that allow for expansion of HSCs in as little as six days. Culture media, isolated cell populations and kits are also provided.