118 results about "Primary culture" patented technology

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 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 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 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.

The application discloses a tissue culture and rapid propagation method for Yangmei of Alocasia macrorrhiza and a special combined culture medium thereof, and the method comprises the steps of sterilizing an explant, carrying out primary culture and subculture and proliferation culture in stages, wherein the primary culture is performed by first inoculating a stem segment into an induction culture medium containing 6-BA 0.8-1.2 mg / L, IBA 0.05-0.15 mg / L and PVP-40 to induce germination, and then transferring the stem segment into an elongation culture medium containing 6-BA 0.1-0.5 mg / L, NAA 0.01-0.05 mg / L and PVP-40 to promote elongation; the new shoots are then transferred into a subculture medium containing 6-BA 1.0-1.8 mg / L, NAA 0.1-0.2 mg / L, GA3 0.5-1.5 mg / L and PVP-40 to proliferate. The application effectively solves the technical problems that Yangmei is prone to brown and death, and it is difficult to simultaneously achieve budding and high growth by means of two-stage hormone stepwise regulation and cooperation with a brown prevention agent, and realizes efficient and large-scale breeding of high-quality seedlings.

The invention discloses a method for separating, culturing and identifying porcine muscle-derived mesenchymal stem cells and fat progenitor cells. The method mainly comprises the following steps: dissociating pig muscle tissues through a tissue processor, obtaining mesenchymal stem cells and adipose progenitor cells by utilizing a flow cytometry sorting technology, and completing in-vitro culture and identification of the cells. According to the technical scheme provided by the invention, the technical problems of primary culture, separation and identification of the mesenchymal stem cells and the adipose progenitor cells of the porcine muscle tissue are effectively solved, and a reliable method is provided for further application of a flow cytometry sorting technology and a myogenic mesenchymal stem cell and adipose progenitor cell primary model in animal nutrition research.

The invention discloses a method for propagating robust artificial seed stems of pinellia ternate through low-cost tissue culture. The method comprises the steps of germination, explant disinfection, establishment of a primary culture system, petiole subculture propagation, tuber subculture propagation and artificial seed stem harvesting. The plant regeneration approach designed by the invention is to induce one end of the petiole to directly generate a small tuber with a complete form under hormone-free concentration, and the small tuber is very similar to the part and the form of the bulbils generated under natural conditions. Tubers are not subjected to a callus stage, are uniform in size and high in hereditary stability, can maintain the characteristics of excellent varieties, and are an optimal way for in-vitro rapid propagation of pinellia ternate. A large number of mature, full, neat and consistent third-generation tubers with high germination rate, stress resistance and storage resistance can be obtained in a short time, the tubers can be directly transplanted to a field without special domestication, the survival rate is high, practical operation and large-scale production are facilitated, and production is not limited by seasons.

The invention relates to the technical field of high-yield collagen MSC (mesenchymal stem cell) engineering construction, and particularly discloses a fixed-point controllable high-yield collagen MSC engineering construction method, which comprises the following steps: 1, MSC separation and primary culture: separating from mammal bone marrow, umbilical cord or adipose tissue to obtain primary MSC, adopting a DMEM / F12 culture medium containing 13% of fetal calf serum, and carrying out primary culture on the primary MSC; carrying out primary culture and passage in a culture environment of 37 DEG C and 5% CO, and screening the fourth generation of MSC in the logarithmic phase as an engineered starting cell; and 2, constructing a collagen synthesis related gene fixed-point editing carrier: designing a fixed-point editing element aiming at a collagen synthesis key gene and an MSC differentiation regulation gene. According to the site-specific controllable high-yield collagen MSC engineering construction method, the collagen expression quantity of the engineered MSC can be greatly improved, the directional synthesis proportion of the type I collagen and the type III collagen is improved, and the requirement of industrialization for high-yield and high-purity collagen is effectively met.

The invention relates to a lung fibroblast culture method, and belongs to the technical field of cell culture. Comprising the following steps: S1, organization preparation; s2, tissue treatment; s3, primary culture: the treated tissue blocks are attached to a culture dish, a culture solution is added, the culture solution is a primary culture system, and culture is performed in an incubator for several days until the culture dish is full of fibroblasts; s4, subculture: digesting the cells subjected to primary culture by adopting conventional 0.25% trypsin, then adding a culture solution which is a subculture system for subculture, and carrying out subculture to a fourth generation; and S5, cell identification. According to the invention, primary and subculture are respectively purified and amplified by virtue of a double culture system of fibroblasts, so that the purification of target cells is facilitated, the cell quality is good, a low-serum growth additive is used as a nutrient substance, the low-serum growth additive is more easily utilized by the cells, the proliferation of the cells is facilitated, and good proliferation activity and phenotypic stability are still kept when the cells are subcultured to the fourth generation; no obvious differentiation phenomenon occurs.

The invention relates to a method for separating, culturing and purifying human amniotic mesenchymal stem cells. The method comprises the following steps: collecting and pretreating a human amniotic membrane, washing, digesting pancreatin, neutralizing and washing, digesting type II collagenase, performing primary culture, performing passage purification and the like. Compared with the prior art, in the pancreatin digestion stage, by increasing the number of times in the pancreatin digestion process, mixing of hybrid cells can be greatly reduced, meanwhile, a large number of stem cells with good activity are obtained, and the extraction success rate of the stem cells is remarkably increased; due to the optimized operation process, tedious steps are reduced, and operability is improved; through multiple purification steps, it is ensured that the finally obtained cells have high purity and a good state.

1. Name of the design product: bioreactor. 2. Use of the design product: the product is an integrated bioreactor, which can complete cell culture (primary culture), expansion culture, cell culture from tissues, medium replacement and cell harvesting. 3. Design points of the design product: in shape. 4. Picture or photo best indicating the design points: front view.

The application discloses a guayule propagation culture medium and a tissue culture rapid propagation method thereof, and belongs to the technical field of tissue culture, and the culture medium comprises MS+0.07 mg / L 6-BA+1 g / L activated carbon+2 g / L agar. The method comprises the following steps: primary induction culture: stem segment explants are induced into seedlings; propagation culture: tissue culture seedlings obtained through the primary culture are selected, stems of axillary buds are taken, and subculture propagation culture is carried out to obtain multiple shoots; rooting culture: the multiple shoots are cultured to obtain rooted tissue culture seedlings; and transplanting: the rooted tissue culture seedlings are transplanted to obtain seedlings. The application can solve the technical problems that the subculture propagation process is extremely easy to vitrify and the propagation coefficient is low, can solve the problems that the excellent seed source of guayule is stable in property and rapid propagation, and realizes large-scale production of high-quality and non-toxic guayule seedlings.

The invention discloses application of keratin in preparation of a bone repair material, and relates to the technical field of biomedical engineering. The preparation method comprises the following steps: extracting bone marrow mesenchymal stem cells, and carrying out osteogenic induction culture under the intervention of keratin with the concentration of 100-800 [mu] g / ml; the culture conditions of the BMSCs are as follows: in a constant-temperature incubator with the temperature of 37 DEG C and 5% CO2, a culture medium contains a precooled complete culture medium; the first liquid changing time of the primary culture of the BMSCs is 48-72 hours, then the liquid is changed once every 2-3 days, and passage is carried out when the cell fusion degree reaches 80%-90%. Keratin is adopted as a natural biological material, has good biocompatibility and biodegradability and can well interact with cells, and immunological rejection is reduced.

The present invention relates to a method for promoting wound healing in a subject suffering from Ectodermal dysplasia in need thereof comprising a step of administering subcutaneously, intradermally or topically to said subject a therapeutically effective amount of a compound which restores the activity of p63. Inventors have performed a primary culture of patient keratinocytes suffering from ectodermal dysplasias with two compounds which restore the activity of p63 (e.g. STIMA-1 and / or PRIMA-1Met). They have shown that there is an important differentiation of the keratinocytes of said patient compared to the cells not treated with these compounds. They observed that the activity of p63 mutated is restored, thus the proliferation and differentiation of keratinocytes from the patient are activated. Moreover, inventors have used PRIMA-1Met by topical application on a young patient suffering from ectodermal dysplasias and shown that said patient presents an improvement on her hand. Typically, severe skin erosions (on hands and feet) are healing when PRIMA-1Met is administered topically on the hand.

The application relates to a method for separating and primary culturing of monopterus albus liver cells. The method comprises the following steps: obtaining monopterus albus liver tissue, perfusing the liver tissue with HBSS buffer solution without calcium and magnesium until blood is completely drained; removing the capsule, blood clots and connective tissue after rinsing, and cutting the liver tissue; sequentially using 0.02% EDTA solution, 0.25% trypsin solution and 50 U / mL collagenase IV working solution to perform three-step digestion treatment on the liver tissue fragments; obtaining a cell suspension by blowing and filtering after the digestion is completed, and then performing gradient centrifugal purification of the cells at rotating speeds of 600 rpm, 250 rpm and 80 rpm in sequence; and finally resuspending the cells in DMEM / F12 culture medium containing 10% FBS and inoculating and culturing the cells. The monopterus albus primary liver cells obtained by the method have the advantages of large quantity, high activity and high purity, and provide a reliable in-vitro model for monopterus albus related research.

The invention relates to the technical field of plant tissue culture, in particular to a sterile rapid propagation method of elaeagnus pungens stem tip meristem. The method comprises the following steps: immersing a stem section in a treating agent, and then washing with sterile water; inoculating the stem tip meristem into a primary induction culture medium which does not contain an exogenous antibacterial agent, and culturing to obtain sterile cluster buds; the method comprises the following steps: selecting robust rootless seedlings, and soaking the base parts of the seedlings in a solution containing IBA for pretreatment; then, the seedlings are inoculated into a rooting culture medium without exogenous auxin to be cultured, and complete regenerated plants are obtained; and hardening seedlings and transplanting. According to the invention, the long-acting silver ion / nano-silver composite protective layer is constructed, and the functional synergistic application of the protective layer in the whole process of subsequent primary culture, proliferation and seedling strengthening and rooting induction is realized; technical contradictions between efficient sterilization and explant activity maintenance, long-term sterile culture and endophyte pollution control, and high-quality root system induction and culture link disjunction in a traditional rapid propagation technology are synergistically solved.

The invention discloses a method for screening edible and medicinal fungi with high yield of microbial proteins and target active components and a kit thereof, and belongs to the technical field of microbial screening. The method comprises the following steps: carrying out primary culture screening on a culture medium containing a specific precursor; performing safe primary screening; carrying out miniaturized culture and rapid protein detection on the safe strains to evaluate the protein potential, and meanwhile, carrying out visual activity screening by utilizing a report system based on functional characteristics of target active ingredients; and finally, carrying out performance verification on the optimized strain. According to the method disclosed by the invention, high-throughput and synchronous evaluation of the protein yield and the active component synthesis potential of the strain is realized, the bottlenecks of low throughput and single target of a traditional method are solved, the excellent edible and medicinal fungus strain with double effects of one strain can be quickly screened, and a core strain screening tool is provided for efficient production of sustainable protein and natural active components.

The invention relates to the technical field of plant cultivation, in particular to a method for establishing a polygonum runcinatum high-frequency regeneration system. The method comprises the following steps: S1, preparing and disinfecting explants, namely disinfecting polygonum runcinatum seeds, inoculating the polygonum runcinatum seeds to a primary culture medium under a sterile condition, and culturing to obtain sterile seedlings; s2, cluster bud induction: cutting terminal buds of the sterile seedlings obtained in the step S1 as explants, inoculating the explants into a first induction culture medium, and inducing the terminal buds to generate cluster buds; s3, proliferation of the cluster buds: after the cluster buds in the step S2 are segmented, the cluster buds are transferred to a second induction culture medium, and proliferation and growth of the cluster buds are promoted; s4, rooting culture: transferring the robust cluster bud individuals obtained by proliferation in the step S3 into a rooting culture medium, and inducing formation of adventitious roots; s5, seedling hardening and transplanting: carrying out seedling hardening treatment on the tissue culture seedlings with completely developed root systems in the step S4, and transplanting the tissue culture seedlings into a culture medium to obtain polygonum runcinatum regenerated plants. The invention establishes a rapid propagation system for polygonum runcinatum.

This invention belongs to the field of biology and discloses a method for co-culturing enteric glial cells (EGCs) and intestinal organoids. Primary cultured and purified enteric glial cells are digested and seeded onto the membrane of a Transwell chamber. Intestinal organoids are placed in individual Matrigel drops in a co-culture plate. The Transwell chambers are placed in the co-culture plate, and growth medium is added to ensure free exchange of the growth medium through the Transwell membrane. The EGC-organoid co-culture system obtained based on this method provides a model for studying diarrhea and intestinal health in large mammals. It also lays the research foundation for developing innovative intervention strategies targeting the enteric nervous system and provides a research model for further exploring the regulatory roles of nutrients or related genes in intestinal health.