Disclosed are methods of releasing or enriching for steviol glycosides produced by yeast. The disclosed methods enhance or improve the release or enrichment of the steviol glycosides.

A method for elongating telomeres of cells comprises steps of: providing physical stimulation directly or indirectly to cells; and culturing a mixture of the cells and a medium for a predetermined time, wherein providing the stimulation directly to the cells comprises applying physical stimulation to the medium containing the cells, and providing the stimulation indirectly to the cells comprises applying physical stimulation to the medium not containing the cells and then mixing the medium and the cells. The method for elongating telomeres of cells is simpler than a conventional method and is superior in terms of time, cost, efficiency, and safety. In addition, the method induces cell division and provides an anti-aging effect, in addition to simply elongating telomeres. Thereby, it is expected that the method can ameliorate and prevent not only problems caused by shortening of telomeres, but also various aging-related diseases and conditions.

The present invention provides a method for preparing 3D brain organoids, comprising the following steps: neurospheres obtained by the RONA method are dissociated into single cells by accutase, plated on a cell culture plate after being counted, cultured in medium A until day 7; neurospheres are cultured in medium B until day 25˜35, and then they are encapsulated by Matrigel; neurospheres are further cultured in media B until day 55˜65, and then they are encapsulated by Matrigel for the second time and cultured continually afterwards. The present invention also provides a medium for culturing 3D brain organoids. The present invention begins with highly purified neurospheres obtained by the RONA method, and neuronal stem cells can be controlled and cultured to achieve true 3D brain organoids with uniform size and structure by this relatively simple method. The 3D brain organoids have six-layered cortical structure of the brain and various subtypes of inhibitory interneuron cells, which are suitable for disease research in vitro, drug screening, etc., and are of great significance in industrialization.

The present invention provides improved and/or shortened methods for expanding TILs and producing therapeutic populations of TILs, including novel methods for expanding TIL populations in a closed system that lead to improved efficacy, improved phenotype, and increased metabolic health of the TILs in a shorter time period, while allowing for reduced microbial contamination as well as decreased costs. The methods may comprise gene-editing at least a portion of the TILs to enhance their therapeutic efficacy. Such TILs find use in therapeutic treatment regimens.

Disclosed is a method for the dissociation of cells. Cells are processed under varying conditions of pH, temperature, and shear to thereby produce different cell products. In one form, the cells are jet cooked at a lower temperature and/or pressure to provide products that are relatively delicate. The remaining cell components may then be subsequently jet cooked under higher temperature and/or shear conditions to provide products that are relatively more robust. Generally, the cells become dissociated, whereby at least one separate cell wall component is substantially separate from the dissociated cell walls.

Cryopreservation of endothelial cell monolayers is one of the major challenges in the cryopreservation of complex tissues. Human umbilical vein endothelial cells (HUVECs) in suspension are available commercially and recently their post-thaw cell membrane integrity was significantly improved by cryopreservation in 5% dimethyl sulfoxide (DMSO) and 6% hydroxyethyl starch (HES). However, cryopreservation of cells in monolayers has been elusive. The exact mechanisms of damage during cell monolayer cryopreservation are still under investigation. Here, we show that a combination of different factors contribute to significant progress in cryopreservation of cell monolayers. The addition of 2% chondroitin sulfate to 5% DMSO and 6% HES and cooling at 0.2 or 1° C./min led to high membrane integrity (97.3±3.2%) immediately after thaw when HUVECs were cultured on a substrate with a coefficient of thermal expansion similar to that of ice. The optimized cryopreservation protocol was applied to monolayers of primary porcine corneal endothelial cells, and resulted in high post-thaw viability (95.9±3.7% membrane integrity) with metabolic activity 12 hours post-thaw comparable to unfrozen control.

The invention relates to a new method for in vitro expansion of CD4.sup.+CD25.sup.HighCD127.sup.−/LowFoxP3.sup.+ Tregs, wherein the process of Treg expansion takes place permanently or temporarily at a temperature below 37° C., optimally at a temperature of 33° C., the isolated Tregs are expanded in SCGM or X-vivo-20 medium supplemented with human serum or with foetal bovine serum, and magnetic beads coated with anti-CD3 and anti-CD28 antibodies at 1:1 (cell:bead) ratio and interleukin-2 are added to the culture.

This disclosure provides a novel method of controlling the glycosylation profile of a protein during production. The disclosure also provides a novel method of improving protein yield while controlling the glycosylation profile of a protein.

The present invention relates to a device and a method for inducing pluripotent cells using energy and, more specifically, has an effect of inducing new type pluripotent cells having pluripotent characteristics by applying energy such as ultrasonic waves, lasers or heat treatment to differentiated cells.

The present application discloses a high-efficiency petroleum degrading bacterium TDYN1.sup.T and use thereof, identified as Falsochrobactrum sp., deposited in the China General Microbiological Culture Collection Center, with a deposit number CGMCC No. 18061. The high-efficiency petroleum degrading bacterium TDYN1.sup.T of the present application is used in petroleum-contaminated soil for degrading, having a good effect, in particular, having a high degradation efficiency when the petroleum is degraded under the conditions of 30±1° C. and pH 7-7.4.