219 results about "Primary culture" patented technology

The invention relates to a marine ranching digital construction method and system based on artificial intelligence, and the method comprises the steps: monitoring plankton in a marine ranching in real time, obtaining a water body image through an image recognition technology, and building a dynamic database, so as to regularly store and update the related data of the plankton. Based on monitoring data, a water quality evaluation model is established, and the correlation among plankton species, density and water quality parameters is analyzed. Factors such as water flow velocity and flow direction are integrated, a flow change model is established, and the influence of water flow on plankton is judged. A primary culture optimization scheme is formulated by simulating the interaction between cultured organisms and planktons, and the culture trend is predicted in real time and the management strategy is adjusted by integrating the water quality evaluation and the output result of the flow change model. According to the scheme, the intelligent management level of the marine ranching is effectively improved, dynamic and scientific management of the culture environment is achieved, powerful support is provided for healthy growth of cultured organisms, and sustainable development of the marine ranching is promoted.

The invention relates to the technical field of microalgae heterotrophic culture and biomass proliferation, in particular to a chlorella pyrenoidosa heterotrophic high-density proliferation method. The specific technical scheme is as follows: the heterotrophic high-density proliferation method of chlorella pyrenoidosa comprises the following steps: pre-culture and heterotrophic propagation of chlorella pyrenoidosa: inoculating a chlorella pyrenoidosa solution into a BG11 culture medium, and culturing for 3-4 days in a constant-temperature shaking table with the temperature of 25 + / -1 DEG C, the rotating speed of 200rpm and the light-dark ratio of 0h: 24h to obtain a primary culture chlorella solution; inoculating the primary culture algae liquid into a pre-sterilized and optimized heterotrophic culture medium, and carrying out expanding culture under the same culture condition for 3-4 days to obtain a secondary expanding culture algae liquid; (2) a heterotrophic high-density culture stage: regularly supplementing a nutrient culture medium containing high-concentration glucose, nitrate and phosphate by adopting an index fed-batch feeding strategy; other conditions are kept consistent during culture; and (3) an in-situ protein accumulation induction stage: when the chlorella pyrenoidosa reaches an exponential growth end stage, adding paclobutrazol into the system, and keeping other conditions consistent while keeping exponential fed-batch culture. According to the method disclosed by the invention, the biomass of the chlorella pyrenoidosa is increased by about 128.6% compared with that of the traditional batch culture, the protein content can reach 49.14%, meanwhile, the configuration of essential amino acids is improved, and the requirements of high-end biomass resource application are met. The method has the advantages of simplicity and convenience in operation and high production efficiency, and is suitable for the fields of high-quality protein extraction and related biomass production.

The invention provides application of a bacteriostatic agent in open type tissue culture and rapid propagation of polyploidy bamboo reeds, and relates to the technical field of tissue culture and rapid propagation of plants, the bacteriostatic agent is applied to the open type tissue culture and rapid propagation process of the polyploidy bamboo reeds, and the use method is that the bacteriostatic agent is added into a culture medium for use or sprayed or oscillated and flushed for use, and the bacteriostatic agent is applied to the open type tissue culture and rapid propagation process of the polyploidy bamboo reeds. The bacteriostatic agent is formed by mixing isothiazolinone and sodium dehydroacetate, the mass addition ratio of the isothiazolinone to the sodium dehydroacetate is (8-5): (2-5), and the method comprises the following steps: S1, primary culture of explants; s2, multiplication culture of seedlings; s3, rooting culture, wherein each liter of the culture medium contains 0.05-1g of the bacteriostatic agent in the above steps. The method has the beneficial effects that under the condition that the method does not need to be controlled in a sterile environment and does not depend on the quality of an inoculator, a large amount of propagation of the polyploidy bamboo reed tissue culture seedlings is realized by adopting an open tissue culture technical mode, the productivity speed of a tissue culture center is improved, and the construction threshold and cost of the tissue culture center are reduced.

A method for separating an exosome from a heterologous expression cell or a primary culture cell, and a use of an exosome in biological analysis, relating to the technical field of biomedicine. The method for separating the exosome from the heterologous expression cell or the primary culture cell of the present invention comprises: carrying out differential centrifugation on a solution to be extracted that contains an exosome, then taking out a first supernatant, carrying out high-speed centrifugation again, respectively adding iodixanol into an obtained second supernatant and a precipitate, and carrying out spontaneous density gradient centrifugation to obtain a separated purified exosome. According to the present invention, an iodixanol spontaneous density gradient centrifugation method is innovatively used to successfully carry out separation and purification to obtain the exosome having high purity and high yield, and the separated exosome is used as an experimental development universal platform for researching a membrane protein structure and functional characteristics in an original ecological cell membrane environment, and is used for biological analysis, drug screening, and disease diagnosis and treatment.

The invention discloses a dental sac mesenchymal stem cell with high periostin expression as well as an extraction and culture method and application thereof, and belongs to the technical field of biomedicine. The dental sac mesenchymal stem cell with high periostin expression is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: C2024384; the extraction and culture method disclosed by the invention comprises the following steps: S1, extraction: cleaning, cutting, digesting and centrifuging dental sac tissues, resuspending, centrifuging and precipitating by using a primary culture medium, culturing until cells are fused, digesting, and carrying out secondary centrifugation and precipitating to obtain primary dental sac mesenchymal stem cells; s2, subculture: resuspending the dental sac mesenchymal stem cells in the step S1 by using a primary culture medium, supplementing a subculture medium, and then subculturing. The invention also discloses the dental sac mesenchymal stem cell with high periostin expression and application of the dental sac mesenchymal stem cell in preparation of medicines for preventing or / and treating periodontal diseases. The method disclosed by the invention has an unexpected effect of enhancing periostin expression of the dental sac mesenchymal stem cells.

The invention relates to the field of plant tissue culture, and discloses a method for culturing tissue culture seedlings of small angel viridae, which comprises the following steps: S1, explant selection and primary culture; s2, carrying out subculture multiplication culture; s3, rooting culture; s4, domesticating and transplanting; in the primary culture stage, MS is used as a basic culture medium, a proper amount of 6-BA, NAA, sucrose and agar are added, and induction of cluster buds is promoted; in the subculture multiplication culture stage, an optimal multiplication culture medium is screened through an orthogonal test, so that the multiplication coefficient is improved; in the rooting stage, key factors such as inorganic salt concentration, NAA and banana puree in MS are optimized, and the number and quality of root systems are improved; in the domestication transplanting stage, the optimal proportion of river sand, coco coir and peat soil is researched, and the transplanting survival rate and the tissue culture seedling quality are effectively improved. By means of the method, efficient propagation of the small angel vitis vinifera and large-scale production of high-quality seedlings are achieved, and important technical support is provided for industrial popularization of the plant.

The invention discloses a tissue culture and rapid propagation method for a fig male tree germplasm material, and belongs to the technical field of plant tissue culture and rapid propagation. The specific method comprises the following steps: (1) obtaining and disinfecting an explant; (2) primary culture of buds; (3) subculture multiplication of buds; (3) rooting culture; and (4) hardening seedlings and transplanting. According to the method, the optimal culture medium for tissue culture and rapid propagation of the fig male tree is obtained by combining different hormones and adjusting different hormone concentration ratios, and the differentiation and proliferation of buds of the fig male tree and the growth and rooting rate of seedlings are remarkably improved. In the explant treatment, tebuconazole carbendazim, 75% alcohol and sodium hypochlorite are adopted for the first time, so that the use of a highly toxic reagent mercury bichloride is avoided; cefotaxime sodium is added into a primary culture medium, so that the pollution rate of explants is reduced; the method is simple to operate, time-saving, labor-saving, safe and environment-friendly, and improves the inoculation quality and efficiency. The method is not limited by seasons, enlarges the excellent male tree population of the figs in a short time, and lays a foundation for cultivating new germplasm of the figs.

The invention discloses a method for extracting 3 beta-acetoxy-15 alpha, 22-dihydroxyhopane from metarhizium anisopliae, which comprises the following steps: firstly, performing activated culture on the metarhizium anisopliae in a PDA (potato dextrose agar) culture medium, then transferring the metarhizium anisopliae into a PDB (potato dextrose agar) culture medium for primary culture, then inoculating the metarhizium anisopliae into a rice culture medium for secondary culture, performing ultrasonic extraction in combination with ethyl acetate extraction and concentration to obtain a crude extract, and performing freeze-drying to obtain the 3 beta-acetoxy-15 alpha, 22-dihydroxyhopane. The crude extract is subjected to separation and purification through a reduced pressure column and a normal phase silica gel column in sequence, and therefore the 3 beta-acetoxy-15 alpha, 22-dihydroxyhopane is obtained. According to the invention, the 3 beta-acetoxy-15 alpha, 22-dihydroxyhopane is separated from the Metarhizium anisopliae for the first time, and it is proved that the 3 beta-acetoxy-15 alpha, 22-dihydroxyhopane can be used for biological control of tea geometrid.

The invention relates to a method for forming a hair follicle-like structure by co-culturing an immortalized hair papilla cell line derived from human hair follicles and keratinocytes. Specifically disclosed is a hair papilla cell line (SV40T-hTERT DPC) which is immortalized by forced expression of SV40T, which deactivates a cancer suppressor gene, in human hair follicle-derived hair papilla cells and hTERT gene, which is a telomerase elongase promoting gene, and keratinocytes, are co-cultured three-dimensionally. From the result, it has been confirmed that a hair follicle-like structure is formed in which a cuticle grows so that a hair follicle analogue in combination with a primary-cuticle hair follicle-derived cell has a shape similar to that of the primary-cuticle hair follicle-derived cell. Therefore, the method can be applied to multiple fields of animal replacement experiments, efficacy evaluation of materials for preventing alopecia / promoting hair follicle growth and the like. Through immortalized hair papilla cells, higher result consistency can be ensured compared with that in the prior art, and stable hair follicle-derived cell supply can be realized, so that the problem of high entry barriers of efficacy evaluation of alopecia materials based on cells is expected to be solved.

The invention discloses a rectal cancer organoid culture kit and a culture method thereof, and particularly relates to the technical field of genetic engineering.The kit comprises a culture medium with specific components, a sample treatment reagent, a passage reagent and a preservation reagent, and the culture medium contains Advanced DMEM / F-12 and multiple factors; the sample treatment reagent can effectively treat tissues; a passage reagent guarantees passage and cryopreservation; and the preservation reagent ensures the cell cryopreservation and resuscitation effects. The culture method covers scaffold and scaffold-free culture processes and comprises the steps of primary culture, subculture, recovery and the like, and the steps are operated finely. The method has the advantages of obvious advantages, fast tissue dissociation, small damage, good cell suspension dispersion, no DMSO toxicity, simple culture, high amplification efficiency, stable passage, wide applicability and high cost performance, and can provide a powerful tool for rectal cancer research, drug screening and the like.

The invention provides a method for inducing test tube tuberous roots under dahlia pinnata tissue culture conditions, which belongs to the technical field of plant tissue culture and comprises the following steps: step 1, obtaining sterile explants; step 2, primary culture: removing a color-changing part on the wound surface of the to-be-inoculated explant obtained in the step 1, and then inoculating the to-be-inoculated explant into a primary culture medium for culture to obtain a sterile test-tube plantlet; step 3, subculture multiplication culture: cutting the seedlings into cluster blocks, and inoculating the cluster blocks into a subculture multiplication culture medium to obtain multiplication test-tube seedlings; step 4, rooting culture; performing illumination culture in a rooting culture medium for 4 weeks to obtain rooted test-tube plantlets; 5, test tube root tuber induction: inoculating the rooted test tube plantlet into a root tuber induction culture medium for illumination culture for 4 weeks to obtain dahlia pinnata test tube sterile root tubers; through segmented induction and addition of different exogenous hormones, the breeding efficiency of dahlia pinnata can be effectively improved, and the breeding period is shortened.

The invention relates to an extraction and culture method of urine-derived stem cells, and belongs to the field of cell biology and regenerative medicine. The method comprises the following steps: coating a culture plate with a gelatin solution; collecting clean midstream urine of healthy people and adding antibiotics; centrifugally separating the cell precipitate; washing with a phosphate buffer solution; re-suspending the precipitate by using a urine stem cell complete culture medium for primary culture; and after the cells grow to a certain fusion degree, carrying out passage amplification. The urine stem cell complete medium comprises a basic medium, fetal calf serum, epidermal growth factors, platelet-derived growth factors and basic fibroblast growth factors. By adopting the method disclosed by the invention, the urine-derived stem cells can be efficiently and stably separated from urine, and the obtained cells have high multiplication capacity, good stemness maintaining capacity, multidirectional differentiation potential and immunoregulation function, and can provide an ideal seed cell source for treating diseases such as diabetic erectile dysfunction.

The invention relates to the technical field of aquatic animal cell culture, in particular to a rapid enrichment and separation method for gonad somatic cells of plectropomus leopardus. Comprising the following steps: selecting healthy plectropomus leopardus for anesthesia; the method comprises the following steps: taking gonad tissues, performing sterile treatment, cutting, centrifugal washing and filtering, inoculating the gonad tissues into a culture medium for primary culture to obtain gonad cells emigrated from tissue blocks, performing trypsin-EDTA digestion and centrifugal washing, and treating the gonad cells with 1 * PBS containing polysorbate; after closing, antibody incubation and washing, the marking effect is verified through a fluorescence microscope, and specific enrichment of gonad somatic cells is achieved; and filtering and purifying the enriched fluorescence-labeled cells, placing the cells under a fluorescence microscope, accurately picking fluorescence-positive cells under a fluorescence visual field, transferring the fluorescence-positive cells into a culture dish containing a complete medium, inoculating the separated fluorescence-positive cells into the optimized complete medium, and carrying out purification culture, so that the cells grow in a wall-attached manner and stable passage is realized.

The application discloses a construction method and application of an ovary cell line of Quasipaa spinosa. The method comprises the following steps: selecting metamorphosis period larva ovary tissue, disinfecting and obtaining the ovary tissue, and then using collagenase I digestion solution to digest the ovary tissue at 26 DEG C for 1 hour; after the tissue block adheres to the wall, using a low-osmotic-pressure complete culture medium to carry out primary culture at 25-27 DEG C, wherein the culture medium is 60% diluted DMEM / F12 culture medium containing 10%-15% fetal bovine serum; when the cell confluence degree is higher than 70%, using trypsin-EDTA to digest the cells for 1-2 minutes to subculture, and the subculture culture medium contains 30% cell conditional culture medium; and finally, the cells are cryopreserved through programmed cooling. The ovary epithelial cell line of Quasipaa spinosa is successfully established for the first time, the survival rate of the cells after recovery is high, and the karyotype is stable. The constructed cell line can be used for amphibian reproductive biology research, compound screening and in-vitro conservation of germplasm resources.

The present disclosure provides a method for in vitro expansion of cryopreserved cord blood-derived regulatory T cells (Tregs) with a high recovery rate, including the following steps: recovering the cryopreserved cord blood-derived Tregs after first expansion, and resuspending the cells with a serum-free medium; and adding a resulting suspension to an expansion culture medium, and conducting a second expansion culture, where a primary culture is conducted for 1 d to 2 d, then a subculture is conducted once every 1 d to 3 d, and a total culture time is 13 d or more. In the expansion culture medium, interleukin-2 (IL-2) is further added. The method of the present disclosure can achieve the second recovery and expansion of cryopreserved Tregs produced after the first expansion. Tregs produced after the second recovery and expansion can have a viability of 90% or more, and can be further expanded (such as third expansion and fourth expansion). The expanded cells can also be used for clinical treatment. Therefore, the method of the present disclosure greatly shortens the time for clinical patients to wait for Tregs, and can meet the clinical on-demand access requirement.

The invention discloses a culture medium composition and a culture method for in-vitro rapid propagation culture of hippeastrum hippeastrum, and belongs to the technical field of plant tissue culture. The adenine, the rhizoma polygonati and the wormwood extract are added into the hippeastrum striatum in-vitro rapid propagation culture medium for the first time, and the culture medium combination for hippeastrum striatum in-vitro rapid propagation culture is developed. The hippeastrum hippeastrum in-vitro rapid propagation culture method established on the basis of the culture medium combination can be used for in-vitro rapid propagation culture of different varieties of hippeastrum hippeastrum, the germination rate of primary culture in the hippeastrum hippeastrum in-vitro rapid propagation culture can be increased, and the differentiation coefficient and the seedling formation coefficient in the hippeastrum hippeastrum tissue culture process can be increased. The efficient and stable hippeastrum striatum in-vitro rapid propagation method established by the invention lays a technical foundation for accelerating a hippeastrum striatum breeding process and commercialized production.

The invention belongs to the technical field of sweet potato seedling detoxification, and particularly relates to a sweet potato seedling detoxification rapid propagation method and application, and the sweet potato seedling detoxification rapid propagation method comprises the following steps: carrying out heat treatment culture on sweet potato seedlings at 36-38 DEG C, disinfecting, and taking stem segments with the length of 0.5-1.5 cm as explants; inoculating the explant into a primary culture medium for culturing; after multiple shoots are induced from the explants, inoculating the explants into a subculture medium for culturing; cutting the cultured cluster buds into single buds, inoculating the single buds into a rooting culture medium for culturing, and detecting the detoxification effect after bud seedlings root to obtain detoxified sweet potato tissue culture seedlings. By means of the method, virus detoxification of the sweet potato seedlings can be achieved, the detoxification rate reaches up to 95%, the survival rate reaches 96.7%, the seedling breeding period of 2-3 months is shortened to 45-50 days, the health level and breeding efficiency of the seedlings are improved, and the yield and quality of sweet potatoes are further improved.

The application discloses a culture medium for tissue culture of Limosella australis, which comprises a primary culture medium, a proliferation and differentiation culture medium and a rooting culture medium. The primary culture medium is MS+6BA 0.4-0.6 mg / L+NAA 0.04-0.06 mg / L+AC 0.8-1.2 mg / L+2,4-D 0.04-0.06 mg / L. The proliferation and differentiation culture medium is modified MS+6BA 0.09-0.11 mg / L+NAA 0.04-0.06 mg / L+AC 1.5-2.5 mg / L. The rooting culture medium is 1 / 2MS+IAA 0.09-0.11 mg / L+AC 2.5-3.5 mg / L. The technical scheme has the highest callus induction rate of 86.7%, the highest differentiation rate of 86.7% and the highest rooting rate of 93.3%.

The invention relates to the technical field of seedling raising, and discloses an efficient tissue culture and rapid propagation seedling raising method for xanthoceras sorbifolia bunge, which comprises the following steps: selecting and pretreating explants, selecting current-year robust branches of the xanthoceras sorbifolia bunge, cutting stem segments with axillary buds as the explants, cleaning, trimming and disinfecting the explants, performing surface disinfection on the pretreated explants, and performing tissue culture and rapid propagation seedling raising on the pretreated explants to obtain the xanthoceras sorbifolia bunge. When the Xanthoceras sorbifolia Bunge is subjected to efficient tissue culture and rapid propagation seedling culture, an operation standard considering thorough sterilization and tissue activity protection is established by standardizing an explant selection standard and executing a step-by-step surface disinfection procedure, so that the microbial contamination rate caused by incomplete disinfection of the explant can be reduced, meanwhile, tissue activity damage caused by excessive disinfection is reduced, and the survival rate of the Xanthoceras sorbifolia Bunge is improved. Therefore, sufficient and healthy sterile initial materials are guaranteed to be obtained, the problems of high primary culture pollution rate and induction starting failure are solved, and a reliable foundation is laid for establishment of a whole tissue culture and rapid propagation system.

The invention discloses a mandarin fish medullary tissue cell line and a construction method and application thereof. The invention belongs to the technical field of aquatic organism cells, and discloses a mandarin fish medulla oblongata cell line MFSB, which is preserved in China Center for Type Culture Collection on December 4, 2024, has a preservation number of CCTCC NO: C2024376, and is named as mandarin fish medulla oblongata cell line MFSB. The construction method comprises the steps of primary culture of mandarin fish medulla oblongata tissue cells, subculture, cryopreservation and resuscitation of the cells and the like, and the mandarin fish medulla oblongata cell line MFSB has the advantages of stable subculture, strong cell line division ability, fast growth speed and good stability, can be used as a cell model or an experimental tool to be applied to scientific research and animal genetic breeding, and has wide application prospects. The probe has broad-spectrum sensitivity to mandarin fish infectious spleen and kidney necrosis virus, mandarin fish double-segment RNA virus, mandarin fish frog iridovirus and mandarin fish rhabdovirus, and can be well applied to separation, culture and detection of mandarin fish virus and research and development of vaccines.

The primary culture method of the porcine ovary granular cells comprises the following steps: step 1, taking a fresh porcine ovary, quickly putting the porcine ovary into a Hank's balanced salt solution preheated at 38 DEG C for transportation, repeatedly washing the ovary with the Hank's balanced salt solution to remove blood and impurities, and then extracting follicular fluid with a 5ml syringe under a sterile condition; according to the method, the granular cells can be effectively separated and cultured from the porcine ovary, and the porcine ovary granular cells with higher purity and better activity can be obtained, so that the subsequent cell biology experiment research, such as gene transfection, protein expression analysis and the like, is facilitated; a cell model is provided for researching the physiological functions (such as hormone secretion and follicle development regulation and control) of the porcine ovarian granular cells, the operation is relatively simple, the cost is relatively low, and the used reagents and instruments are common in a cell culture laboratory and are convenient to popularize and apply.

The invention relates to an isolated culture method of functional Chinese softshell turtle ovarian granular cells. The isolated culture method comprises the following steps: S1, collecting ovarian tissues: collecting Chinese softshell turtle ovarian tissues; s2, separating and collecting a granular cell layer; s3, digesting and collecting granular cells; s4, primary culture of granulosa cells: transferring the collected cells into a cell culture bottle containing 4 ml of a complete culture medium for primary culture, wherein the complete culture medium comprises the following main components: a DMEM culture medium, 15% of fetal calf serum, 1% of penicillin streptomycin and 1% of an insulin-transferrin-selenium additive; and S5, subculturing the granular cells. The isolated culture method and the culture medium system provided by the invention are simpler and more time-saving and labor-saving; according to the method, the aromatase CYP19A1 can be expressed normally, the characteristics of the granular cells are kept, the aromatase CYP19A1 can be expressed normally, it is indicated that the in-vitro cultured cells can keep the functionality of the cells, and the in-vitro proliferation capacity of the granular cells is improved to a great extent.

The invention discloses a factory-like efficient breeding method and system for virus-free seed potatoes in a potato bottle, and relates to the technical field of agricultural biology. The method comprises the following steps: selecting healthy buds, stripping stem tips, carrying out primary culture to obtain seedlings, screening virus-free seedlings through RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection, and transferring the virus-free seedlings to a culture flask containing a liquid MS culture medium for expanding propagation; hardening the propagated virus-free seedlings to obtain stress-resistant virus-free seedlings, inoculating the stress-resistant virus-free seedlings into a tissue culture bottle containing an induction culture medium, performing induction culture under set illumination and temperature, and dynamically adjusting a nutrient solution to obtain virus-free seed potatoes; and classifying and screening the detoxified seed potatoes and storing in a specific environment. The system comprises a sterile operation unit, a multi-stage culture unit, a seedling hardening unit and a graded storage unit. The method can efficiently breed the virus-free potato seeds, reduces the virus infection risk, improves the breeding efficiency and the potato seed quality, and is suitable for industrialized production.

The invention discloses a stipa purpurea tissue culture and rapid propagation method and a special culture medium. The method comprises the following steps: (1) selecting and pretreating an explant; (2) primary culture; (3) subculture; (4) rooting culture; and (5) seedling hardening and transplanting. Through an optimized explant disinfection process, a culture medium system based on N6, a specific plant growth regulator combination and addition of enzyme hydrolysis casein in a rooting stage, a breakthrough of a stipa purpurea tissue culture and rapid propagation technology is achieved, the pollution rate is lower than 5%, the multiplication coefficient reaches 4.2 or above, the rooting rate exceeds 93%, the transplanting survival rate reaches up to 89%, and the tissue culture and rapid propagation technology has the advantages that the tissue culture and rapid propagation efficiency is high. The whole rapid propagation period only needs 60-70 days, and efficient and reliable technical support is provided for large-scale and standardized production and ecological restoration application of stipa purpurea.

The invention belongs to the technical field of plant tissue culture, and provides a composite systemic bactericidal solution for lily bulb pretreatment and a matched disinfection and primary culture method thereof. Each liter of the working solution comprises 4-8 g / L of 50% carbendazim wettable powder, 2-6 g / L of 50% chlorobromoisocyanuric acid soluble powder and 1-2 g / L of metalaxyl-hymexazol aqueous solution. The method sequentially comprises the steps of appropriate dehydration of bulbs, soaking in a composite systemic bactericidal solution, standing for absorption, surface sterilization, short soaking in a protective solution and continuous bacteriostatic culture. According to the technical scheme, the limitation of traditional surface sterilization is broken through, deep and surface double sterilization is considered, the multi-mechanism and multi-link synergistic effect is achieved, the pollution rate (smaller than or equal to 3%) of lily bulb primary culture is reduced, mercuric chloride is avoided, and the method is safe, environmentally friendly and suitable for large-scale culture of lilies and other bulb crops.

The invention relates to the technical field of plant tissue culture, in particular to a method for tissue culture and rapid propagation of polygala japonica. Comprising the following steps: sterilizing and disinfecting explants; carrying out primary culture on the sterilized and disinfected explant; carrying out multiplication culture on primary cluster buds; carrying out rooting culture on rootless tissue culture seedlings; domesticating and transplanting rooting tissue culture seedlings. A primary culture medium, a proliferation culture medium and a rooting culture medium are reasonably designed according to different growth regulator formulas by taking a stem section with buds of polygala japonica as an explant material and are respectively used for performing clumpy bud induction, clumpy bud proliferation and rooting induction on the explant material, the total culture process only needs about 100 days, the breeding efficiency is high, and the survival rate is high. The inductivity, the tufted bud multiplication coefficient and the rooting rate of the explant tufted buds all reach a relatively high level, the cultured tissue culture seedlings are good in quality and high in transplanting survival rate, the tissue culture and rapid propagation effects of the polygala japonica are good, and a solid foundation is laid for realizing industrialized production of the polygala japonica.

The invention discloses a siRNA molecule for interfering sheep TBXT gene expression in primary culture of sheep bone marrow mesenchymal stem cells and application of the siRNA molecule, and relates to the technical field of molecules. According to the invention, a method for in-vitro targeting sheep TBXT gene interference siRNA transfected cells is established by synthesizing a specific targeting combination sheep TBXT gene small fragment sequence, and the mRNA expression level of the TBXT gene of sheep bone marrow mesenchymal stem cells can be effectively interfered. A further research proves that the proliferation efficiency of the sheep bone marrow mesenchymal stem cells is obviously reduced after the TBXT gene is interfered. The TBXT gene is helpful for researching the biological function of the TBXT gene participating in embryonic caudal vertebra development regulation, and has good application value for researching the molecular mechanism of the sheep tail type character.

The application discloses a tissue culture propagation method of eriochloa contracta. The creeping stem section of the eriochloa contracta is collected, washed, and then sterilized and dried; the sterilized and treated stem section is inoculated into a bud induction medium of MS+1-1.5 mg / L 6-BA+0.1-0.5 mg / L NAA+30 mg / L sucrose+6.5 mg / L agar to perform bud induction, and after 15-20 days of culture, the adventitious bud seedling is obtained; the adventitious bud of about 1-1.5 cm induced in the primary culture medium is inoculated into a proliferation culture medium to perform subculture, and the next step can be divided into single bud expansion or direct transplanting domestication. The two-step method only needs primary culture-proliferation and rooting to be simultaneously performed, and the transplanting survival rate is high.

Disclosed is a chemically defined airway basal stem cell culture medium. The provided culture medium involves a simple preparation method and high safety, and does not require a feeder layer or a matrigel-coated culture plate. By means of the culture medium, primary cultured airway basal stem cells achieve good stability; and after multiple passages, the normal morphology of the basal stem cells can be maintained and good differentiation potential can be maintained. In addition, the culture medium has clear components, and can meet both scientific research-grade and clinical-grade applications of airway basal stem cells.

The invention discloses an osteoclast culture method as well as a culture medium and application thereof, the culture method is based on primary monocyte extraction, inoculation is performed according to a specific density, then quantitative M-CSF is added, and culture is performed overnight; on the second day, when the cell confluence degree of the mononuclear bone marrow hematopoietic stem cells is 5-30%, supplementing quantitative RANKL into each hole, continuously culturing for 2-4 days, then changing the solution, and obtaining mature osteoclasts on the sixth-seventh day. According to the invention, the optimization relationship among the cell activity, the cell density and the culture medium in the primary culture process of the osteoclast is deeply studied, and from the perspective of cost reduction and efficiency improvement, the use cost of cell factors is remarkably reduced, the number of times of liquid change is reduced, and the traditional culture time of 10-12 days (calculated from cell extraction) is shortened to 6-7 days. The average diameter of the mature osteoclast cultured by the method is larger, the number of fused cell nucleuses is larger, the space proportion of the mature osteoclast is not less than 80%, and the cell quality is obviously higher than that in the prior art.