5 results about "Dispase" patented technology

The invention relates to the technical field of cell biology, and particularly discloses an isolated culture method of primary meibomian gland epithelial cells, the obtained primary meibomian gland epithelial cells and application of the primary meibomian gland epithelial cells. The method comprises the following steps: efficiently and mildly digesting meibomian gland tissues by adopting a compound digestive juice consisting of I-type collagenase, dispersing enzyme and trypsin in a specific ratio; stopping digestion by using a serum-containing neutralizing solution and collecting cells; the method comprises the following steps: promoting cell attachment by using a serum-containing initial culture medium, and carrying out selective amplification by using a serum-free proliferation culture medium to finally obtain high-purity and high-activity primary meibomian gland epithelial cells. The method solves the problems of long digestion time, low cell yield, poor purity and the like in the traditional method. The obtained cell can be used for constructing a meibomian gland dysfunction model, screening drugs and researching lipid metabolism, and an important technical platform and a cell model are provided for research and treatment of related eye diseases.

The invention discloses a kit and a method for preparing alligator sinensis tissue single cells, and belongs to the technical field of single cell sequencing. The kit comprises Collagenase I, DNase I, Dispase and a DMEM (Dulbecco Modified Eagle Medium) culture medium. The kit and the method provided by the invention fill the blank of the Yangtze alligator tissue single cell preparation technology. The kit is simple in component and easy to prepare, the method has very high repeatability and success rate, a large number of high-motility and low-impurity single cells can be obtained, and the kit can be applied to single cell sequencing and more application fields.

The application belongs to the technical field of biological medicine, and particularly relates to a thymus tissue dissociation reagent, a kit and application thereof in obtaining thymus epithelial cells. The thymus tissue dissociation reagent at least comprises collagenase D, dispersinase II, deoxyribonuclease I, dissociation enzyme and a dissolving reagent; and collagenase II and / or collagenase IV and / or trypsin. The cell survival rate of the thymus tissue dissociation reagent after dissociating the thymus tissue is maintained at a high level of 95.7% to 99.6%, which is significantly higher than that of the prior art; and after sorting, the number of thymus epithelial cells enriched and obtained from each gram of thymus tissue is 1.38e 5 ~1.06e 6 , and the success rate of enriching and obtaining thymus epithelial cells is as high as 95%, which is significantly higher than that of the prior art; and the proportion of epithelial stem cells in the thymus epithelial cells is 65.7%, which is significantly higher than that of the prior art.

The invention particularly relates to a human pneumosarcoma-like carcinoma organ as well as a construction method and application thereof, and belongs to the technical field of biological medicines. The organoid is selected from TJLCI-PSC-O-1 with the preservation number of CCTCC (China Center For Type Culture Collection) NO: C2025218 or TJLCI-PSC-O-2 with the preservation number of CCTCC NO: C2025334, or an amplified culture of an epithelial marker CK and a mesenchymal marker VIMENTIN which are stably co-expressed by the organoid. According to the construction method, a collagenase / dispersing enzyme and DNase I combined stepped enzymolysis system and a three-dimensional culture system are adopted, and high yield and long-term stable amplification of cells are realized. The model highly simulates the pulmonary sarcomatoid carcinoma pathological structure and epithelial-mesenchymal transition phenotype for the first time, can be used for in-vitro drug sensitivity test, in-vivo tumor formation model construction and drug screening, and provides a key platform for basic research and precise treatment of diseases.

This invention discloses a kit and method for preparing single cells from reptiles, belonging to the field of single-cell sequencing technology. The kit includes Collagenase I, DNase I, Dispase, and DMEM culture medium. The kit and method of this invention fill the gap in single-cell preparation technology from reptile tissues, especially the Chinese alligator. The kit has simple components and is easy to prepare; the method has high reproducibility and success rate, and can obtain large quantities of single cells with high viability and low impurities, which can be used for single-cell sequencing and other applications.