7results about How to "Achieve multiplication" patented technology

The invention discloses a human-derived iPSC (induced pluripotent stem cell)-induced liver organ and a preparation method thereof. The method comprises the following steps: digesting human induced pluripotent stem cells obtained by culture into single cells; single cells are prepared into cell clusters through self-balling, or cell matrigel microspheres are prepared by using a T-type microfluidic technology and then are distributed one by one, so that one cell cluster is formed in each pore of the low-adsorption 96-pore plate; carrying out step-by-step induction culture on the obtained cell cluster, and culturing in a culture medium I for 5 hours; replacing with a culture medium II and culturing for 2 days; a culture medium III is used for culturing for 2 days; replacing with a culture medium IV and culturing for 5 days; replacing with a culture medium V, and culturing for 6 days; and replacing with a culture medium VI, and culturing for 10 days to obtain the mature human liver organ. According to the invention, liver organs with human normal liver cell characteristics and protein expression can be formed within 25 days, and the method can realize homogenization and high efficiency of iPSC induction.

The invention discloses a zanthoxylum bungeanum tissue culture method, which belongs to the technical field of tissue culture, and comprises the following steps: (1) callus induction: taking zanthoxylum bungeanum tender stems, disinfecting, and carrying out callus induction culture; (2) multiplication culture: inoculating the callus into a multiplication liquid culture medium for oscillation illumination culture, and spraying a dispersing agent in the culture process; (3) adventitious bud induction: carrying out adventitious bud induction culture on the embryogenic callus obtained by multiplication culture; (4) adventitious bud elongation: cutting the induced clumpy buds into small clumpy buds for adventitious bud elongation culture; (5) performing rooting culture: when the adventitious buds extend to 3-4cm, performing single-plant separation on the adventitious bud clusters, and performing rooting culture; and (6) seedling hardening and transplanting: opening the bottle opening, hardening the seedlings for 3-5 days, taking out the rooted seedlings, and transplanting, so that the problem that high differentiation rate, high density, high quality and high survival rate are difficult to achieve at the same time in the pepper tissue culture process is solved.

The invention provides a method for fermenting roxburgh rose pomace by using bacillus subtilis and saccharomycetes, and belongs to the technical field of pomace fermentation. The method specifically comprises the following steps: S1, treating rosa roxburghii tratt fruit residues: cleaning fresh rosa roxburghii tratt fruits, selecting, juicing, reserving the fruit residues, bagging, sealing, and freezing for later use; s2, activation of strains: bacillus subtilis: controlling the pH value of glucose, yeast extract, beef extract and sodium chloride to 7.0-7.5, sterilizing, cooling, and inoculating the strains in a constant-temperature shaking table for culture; yeast: adding a certain amount of yeast powder into warm water to uniformly disperse the yeast powder, adding cane sugar, and activating in a constant-temperature shaking table; s3, pomace fermentation: preparing the roxburgh rose pomace and water according to a certain proportion, inoculating a bacillus subtilis and saccharomycetes strain liquid seed culture medium, performing compound inoculation and culture fermentation according to a certain inoculation proportion, and drying after fermentation is finished to obtain a pomace fermentation product. The fermentation method has the advantages of simple process, safety, no toxicity, short fermentation period, low cost, high efficiency and the like.

The invention provides a method for preparing cellulase and application of the cellulase. The method for preparing the cellulase comprises the following steps: S1) improving the expression level of an SNC1 gene in trichoderma reesei to obtain trichoderma reesei engineering bacteria; s2, the trichoderma reesei engineering bacteria are cultured and collected to obtain the cellulase, the problem that in the prior art, a large amount of cellulase is difficult to produce through trichoderma reesei under the culture condition containing glucose can be solved, and the method is suitable for the technical field of microorganisms.

The invention relates to a single bacterium screening and culturing method based on a digital micro-fluidic system, which comprises the following steps: separating on a chip of the digital micro-fluidic system to obtain single bacterium liquid drops; the method comprises the following steps: injecting a screened substance on a chip of a digital microfluidic system, separating to obtain single-electrode screened substance liquid drops, controlling the single-electrode screened substance liquid drops, mixing with single bacterium liquid drops to obtain mixed liquid drops, setting temperature and time, culturing bacteria in the mixed liquid drops, injecting new bacterium liquid, and repeatedly culturing. According to the present invention, after the single bacterium liquid drops are efficiently obtained with high throughput, the bacteria are screened, and the separated single bacterium liquid drops can be subjected to the in-situ repeated culture on the chip so as to achieve the proliferation of the bacteria without transferring to the outside of the chip, such that the obtained single bacteria cannot grow due to other transfer reasons, and the obtained single bacteria can be easily obtained. Finally, the sample is lost, and the pollution rate is reduced.

This invention discloses a humanized Sirpα The construction methods of immunodeficient mice and their application in human immune reconstitution belong to the field of medical experimental model construction technology. Rag2 ‑ / ‑ IL2rg ‑ / ‑ Using animals with T, B, and NK cell immune dysfunction caused by mutations as the background strain, sgRNA and dsDNA were specifically designed, and their endogenous components were processed using CRISPR / Cas9 gene editing technology. Sirpα The entire genome was modified to be humanized, thus creating a humanized genome. Sirpα Immunodeficient mice. This method is simple to operate, highly reproducible, and constructs a fully humanized mouse. Sirpα Immunodeficient mice exhibit a higher immunodeficient phenotype and can achieve a higher rate of human T cell chimerism after transplantation with human peripheral blood mononuclear cells, while effectively reducing the risk of graft-host disease.

The invention discloses application of a dithiocarbamic acid compound in preparation of a medicine for preventing and / or treating osteoporosis, and belongs to the technical field of medicines. Research finds that the dithiocarbamic acid compound can effectively prevent and / or treat osteoporosis, improve the bone mineral density of osteoporosis patients and reduce the fracture risk. Meanwhile, the research finds that when the dithiocarbamic acid compound and the calcium supplement are combined for medication, the prevention and / or treatment effect on the osteoporosis can be better achieved.