528 results about "Proliferation rate" patented technology

Nucleic acid drug carriers comprise a nucleic acid carrier complexed with a drug, wherein the nucleic acid carrier and the drug are associated non-covalently, and optionally other agents such as spacer, transfection agents, and targeting agents. The nucleic acid drug complex are discovered to have permissive or refractory uptake depending on many factors including cell type, proliferation rate, among others. The refractive uptake of the nucleic acid drug complex are shown to be useful in the nucleic acid targeting of drugs, both in vitro and in vivo. Novel drug compositions are disclosed that effectively reduce the toxicity of drugs while maintaining drug activity and enhancing a drug's therapeutic index.

The invention relates to a ginseng adventitious root induced proliferation method, which comprises the following steps: cutting ginseng tissue culture seedlings into small tissue blocks and then inoculating into a solid induced culture medium for inducing to form adventitious roots, cutting the adventitious roots into small adventitious root blocks and then inoculating into a liquid proliferation culture medium for performing proliferation culture of the adventitious roots, wherein the solid induced culture medium and the liquid proliferation culture medium take 1 / 2 MS(-N) culture medium as basic culture mediums and contain 1-10mg / L indolebutyric acid. By utilizing the method provided by the invention, the quantity of the adventitious roots obtained through induction can be obviously increased, the adventitious roots obtained through induction are subjected to proliferation culture, and compared with the method for performing adventitious root induction by utilizing callus tissues, the proliferation rate of the adventitious roots is higher, and the proliferation effect is better.

The invention discloses a serum-free medium for culturing placenta mesenchymal stem cells. The serum-free medium takes a DMEM (Dulbecco Modified Eagle Medium) culture solution as a basis and also contains a fibroblast growth factor receptor 2, growth hormone, insulin, transferrin, glutathione, BMP-4, L-glutamine, sodium pyruvate, non-essential amino acids and beta-mercaptoethanol. According to various serum-free media provided by the invention, growth and proliferation of the placenta mesenchymal stem cells in a serum-free medium system can be effectively promoted, the placenta mesenchymal stem cells have higher growth and proliferation rate in the serum-free medium system compared with a serum cell culture medium, the characteristics of the stem cells are preserved, the serum-free medium has multiple differential potentials, and the stem cells can be directionally induced into fat cells and osteoblasts.

The disclosed methods and apparatus provide for the automatic segmentation and analysis of the overall migration rate and proliferation rate of cells that may be used with any resolution image and without the need to prepare the sample or the image before the image is analyzed. In particular embodiments, the method or apparatus of analyzing a cell image comprise performing a structure tensor analysis and / or regularization and / or a histogram-based analysis and / or a level-set analysis to classify pixels in the image into a region of interest (ROI), corresponding to cell clusters, and a non-significant region. Methods and apparatus for cell migration analysis comprise means for computing the areas of the segmented ROIs for a set of images. Methods and apparatus for cell proliferation analysis comprise means for counting the number of cells within the segmented ROIs for a set of images.

The invention relates to a tissue culture and rapid propagation method for a pinellia tuber plant and belongs to the technical field of plant cell engineering. Organs such as sterile blades, leaf stalks, cluster buds and the like of pinellia tuber are used as inoculating materials. The method comprises the following steps of: transplanting the inoculating materials into an intermittent submerged culture reactor, and performing two stages of proliferating and inducing, and rooting and generating and culturing tubers; and after the proliferating culture is finished, replacing a proliferation culture medium with a rooting and tuber generating culture medium under the aseptic condition so as to promote the forming of the tubers of the pinellia tuber. By the method, the automation degree in the production process of pinellia tuber seedlings is greatly improved, the human input is reduced during the culture, and the proliferation rate is greatly improved. The pinellia tuber seedlings which are produced by the reactor have the advantages of no pathogenic bacteria, uniform and stable heredity and the like. The survival rate is obviously improved, the labor cost is obviously lowered, and the guarantee for producing a great number of high-quality seedlings and isolated tubers at low cost is provided. By the method, the seedling quality during the plantation of pinellia tuber medicinal materials is improved, the seedling cost is lowered, and economic benefits are obviously improved and the same time.

This invention relates to human adipose tissue-derived multipotent adult stem cells. More particularly, the invention relates to human adipose tissue-derived multipotent stem cells, which can be maintained in an undifferentiated state for a long period of time by forming spheres and have high proliferation rates, as well as methods for isolating and maintaining the adult stem cells, and methods for differentiating the multipotent adult stem cells into nerve cells, fat cells, cartilage cells, osteogenic cells and insulin-releasing pancreatic beta-cells. Also, the invention relates to cellular therapeutic agents for treating osteoarthritis, osteoporosis and diabetes and for forming breast tissue, which contain the differentiated cells or the adult stem cells. Although the multipotent stem cells are adult stem cells, they have the ability to differentiate into osteogenic cells, nerve cells, astrocytes, fat cells, chrondrogenic cells or insulin-releasing pancreatic beta-cells, and so are effective in treating osteoporosis, osteoarthritis, nerve disease, diabetes, etc. Also, the stem cells form spheres in a serum-free medium containing CORM-2, and thus can be maintained in an undifferentiated state for a long period of time. Also, the stem cells have very high proliferation rates. Accordingly, the stem cells are useful as cellular therapeutic agents.

This invention relates to human adipose tissue-derived multipotent adult stem cells. More particularly, the invention relates to human adipose tissue-derived multipotent stem cells, which can be maintained in an undifferentiated state for a long period of time by forming spheres and have high proliferation rates, as well as methods for isolating and maintaining the adult stem cells, and methods for differentiating the multipotent adult stem cells into nerve cells, fat cells, cartilage cells, osteogenic cells and insulin-releasing pancreatic beta-cells. Also, the invention relates to cellular therapeutic agents for treating osteoarthritis, osteoporosis and diabetes and for forming breast tissue, which contain the differentiated cells or the adult stem cells. Although the multipotent stem cells are adult stem cells, they have the ability to differentiate into osteogenic cells, nerve cells, astrocytes, fat cells, chrondrogenic cells or insulin-releasing pancreatic beta-cells, and so are effective in treating osteoporosis, osteoarthritis, nerve disease, diabetes, etc. Also, the stem cells form spheres in a serum-free medium containing CORM-2, and thus can be maintained in an undifferentiated state for a long period of time. Also, the stem cells have very high proliferation rates. Accordingly, the stem cells are useful as cellular therapeutic agents.

The invention discloses a fast tissue culture reproducing method of actinidia eriantha, belonging to the technical field of plant tissue culture. The method comprises the following steps: (1) tissue culture medium preparation; (2) explant selection and sterilization; (3) inducing culture; (4) proliferation culture; (5) rooting culture; (6) tissue culture plantlet acclimatization and transplant, and the like. The fast tissue culture reproducing method has high culture medium pertinence and good applicability, the inductivity of an initial sprout of an explant reaches more than 80 percent, the proliferation rate of a successive sprout reaches 2-7 times, the plantlet height can reach 2-5 centimeters for 20-30 days, the quality of tissue culture plantlets is enhanced, the rooting rate reachesmore than 95 percent, and the transplant survival rate reaches more than 95 percent. The invention can be popularized and applied to fruit tree plantlet enterprises.

The invention discloses a method for the tissue culture of Lycoris haywardii and a rapid propagation technique. The method comprises the following: 1) a step of preparing culture mediums, and components of a basic culture medium and the culture mediums in every stage of tissue culture, as well as the weight of every component contained in each liter are as follows: the basic culture medium comprises MS or 1 / 2MS, wherein the basic culture medium comprises 20 to 40 g / L of sucrose and 8 g / L of agar and has the pH of 5.8; an induction culture medium comprises MS, 0.2 to 1.0 mg / L of TD and 2 g / L of activated carbon; a proliferation culture medium comprises MS, 3 to 7 mg / L of 6-BA and 0.5 to 2.5 mg / L of NAA; a strong seedling culture medium comprises MS, 0.5 to 2.5 mg / L of 6-BA and 0.5 to 1.5 mg / L of NAA; and a rooting culture medium comprises 1 / 2MS, 0.5 to 2.5 mg / L of KT, 0.5 to 2.5 mg / L IBA and 2 g / L of activated carbon; 2) a step of selecting and sterilizing explants; 3) a step of carrying out induction culture, proliferation culture, strong seedling culture and rooting culture; and 4) a step of domesticating and transplanting tissue culture seedlings. The method has the advantages that the induction rate of Lycoris haywardii buds reaches over 90 percent; the proliferation rate of each week is over 500 percent; the growth rate of the Lycoris haywardii buds is accelerated; the browning phenomenon of the explants is effectively prevented; and the rooting of the tissue culture seedlings is promoted at the same time when bulblet browning is prevented.

Human adipose tissue-derived multipotent adult stem cells are provided, which are characterized by the ability to be maintained in an undifferentiated state for a long period of time by forming spheres and which have high proliferation rates. Also provided are methods for isolating and maintaining the adult stem cells, and methods for differentiating the multipotent adult stem cells into nerve cells, fat cells, cartilage cells, osteogenic cells, muscle cells, endothelial cells, hepatic cells and insulin-releasing pancreatic β-cells. Also provided are cellular therapeutic agents for treating osteoarthritis, osteoporosis, nerve disease, diabetes and for forming breast tissue, which contain differentiated cells or the adult stem cells.

A method of detecting prostate tumorigenesis in a subject, the method including the steps of (a) obtaining a sample from the prostate of the human subject, (b) detecting quantitatively or semi-quantitatively in the sample a level of expression for PKC-ι and (c) comparing the expression level in (b) to a level of expression in a normal control, wherein overexpression of PKC-ι, with respect to the control, indicates the presence of prostate cancer in the subject. The present invention is based upon the discovery that PKC-ι levels are elevated during prostate tumorigenesis. Furthermore, the proliferation rate of the tumor correlates with the level of PKC-ι. The invention also provides methods of treating prostate cancer by administering to the subject a compound that inhibits the expression of PKC-ι. The compound can be a small interfering RNA (siRNA) molecule.

The invention provides a serum-free adipose-derived stem cell culture medium, belonging to the technical field of stem cells. The serum-free adipose-derived stem cell culture medium comprises an IMDM (Iscove's modified Dulbecco's medium) basal culture medium, recombinant human insulin, recombinant transferrin, vitamin C, human serum albumin, fibroblast growth factor, platelet-derived growth factor, epidermal growth factor, insuline-like growth factor, transforming growth factor, fiber connexin, fetuin, tea polyphenol and stem cell growth factor. The serum-free adipose-derived stem cell culture medium provided by the invention does not need to perform culture bottle coating, and has the advantages of cell adherent property, favorable morphology and high cell proliferation rate.

The invention provides a serum-free medium for human umbilical cord mesenchymal stem cells and belongs to the technical field of stem cells. The serum-free medium comprises a DMEM basic medium and further comprises recombinant human insulin, human serum albumin, transferrin, fibronectin, vitamin C, biotin, stem cell growth factors and stem cell factors. The serum-free medium for human umbilical cord mesenchymal stem cells provided by the invention needs no enveloping of a culture flask, operating is convenient and simple, cell proliferation rate is high, good stem cell pedomorphism and stem cell characteristics are kept, the serum-free medium has a good potential for cell induced differentiation, and the cost is lowered.

The invention relates to a subsurface wetland system capable of improving nitrogen and phosphorus removal effects of tail water in sewage plant and avoiding bioclogging. The subsurface wetland system comprises a water distribution zone, a purifying zone and a water collection zone which are sequentially arranged, wherein the water distribution zone is connected with a water inlet pipe, the water collection zone is connected with a water outlet pipe and a sulfur / limestone mixed matrix, a gravel matrix and zeolite matrix are sequentially arranged inside the purifying zone according to the water flowing direction from front to rear. Compared with the prior art, the subsurface wetland is combined with sulfur autotrophic denitrification to remove nitrate in the influent water; since thiobacillus denitrificans slowly grows and the proliferation rate of thiobacillus denitrificans is far lower than that of heterotrophic bacteria, when efficient denitrification nitrogen removal is performed in the subsurface wetland system, the clogging of the subsurface wetland system due to the mass proliferation of microbes cannot be caused and meanwhile, limestone in the mixed matrix is adopted to achieve the removal of phosphates in the influent water.

The invention discloses a method for stimulating the rapid proliferation of CIK (Cytokine-induced Killer) cells by using DC (Dendritic Cells). The method comprises the following steps: separating peripheral blood monouclear cells (PBMC); performing induction culture on cytokine-induced killer cells, i.e., performing induction culture on the CIK cells; performing induction culture on the DC; and detecting the phenotype CD3<+>CD56<+> of the CIK cells. According to the method, the numbers of the CIK cells are amplified substantially within a short period while the CD3<+>CD56<+> ratio of the surfaces of the CIK cells is not changed sharply, so that the amplification time of the CIK is shortened obviously; and the proliferation rate is improved at the same time. Therefore, the important scientific research value is obtained.

The invention discloses a human serum-free culture medium and a preparation method thereof. The human serum-free culture medium uses eleven raw materials, namely human serum albumin solution for treatment, human recombinant insulin solution, human transferrin solution, human cholesterol solution, human catalase solution, 2-mercaptoethanol solution, ascorbic acid solution, linoleic acid solution, ethanolamine solution, human vitronectin solution and L-glutamine solution; the added protein and lipid are both from the blood plasma, serum or tissue of human, the protein is pharmaceutical-grade orhighly purified human protein or human recombinant protein, the protein and lipid do not contains any animal component, the other components all meet the United States Pharmacopoeia or national standards; and the human serum-free culture medium is qualified through the cell culture test and is clinical, safe and reasonable human serum-free culture medium. The proliferation rate of the CIK cell cultured and inducted by the human serum-free culture medium is better than that of the CIK cell cultured and inducted by the culture medium with serum, the cell CD3+CD56+ percentage and the killing rate to the K562 leukemic cell are similar to that of the culture medium with serum. By adopting the human serum-free culture medium, the safety and standardization of cell therapy can be increased.

The invention discloses a method for cultivating a subcultured bud of an upright crown tissue culture seedling of fir wood. The method comprises the following steps: an excellent clonal annual burgeon of the fir wood is used as an explant, a breeding bud is obtained through disinfection, sterilization and tissue culture, the breeding bud is innoculated in a proliferation culture medium comprising 1 / 2MS, 1.0mg / L of 6-BA and 0.5mg / L of IBA for inducing the cluster buds to sprout, the subcultured bud which is germinated at the base of the breeding bud and is healthy in growth is selected and is cut in a common rooting culture medium, seedlings are exercised and transplanted, and when the height of the seedlings of the fir wood is 15-30 cm, the seedlings are taken out of a garden for forestation. When the method is used, the proliferation rate of the subcultured bud of the fir wood is as high as 4-6, the breeding speed is high, the seedling culturing number is large, the tissue culture subcultured bud with high genetic stability, 100% of the upright crown rate of the tissue culture seedling and more than 96% of the rooting rate can be in batch production in a short period, and the requirements of the construction and the development of a fir wood reserving base of China for good seedlings can be met, so that the method has better economic benefits, social benefits and ecological benefits.

The invention provides a serum-free human amniotic mesenchymal stem cell culture medium, and belongs to the technical field of stem cells. The serum-free human amniotic mesenchymal stem cell culture medium comprises the following components: an IMDM basal culture medium, recombinant human insulin, recombinant transferrin, vitamin C, human serum albumin, fibroblast growth factors, platelet-derived growth factors, epidermal growth factors, insulin-like growth factors, transforming growth factors, fibronectin, fetuin, putrescine salt and recombinant human activin. The serum-free human amniotic mesenchymal stem cell culture medium provided by the invention does not need a culture bottle coating process, and is good in cell adherence and morphology and high in cell proliferation rate.

The present invention provides cyclic RNA circ-ZKSCAN1 use, the objective existence of circ-ZKSCAN1 gene is first confirmed; expression status of the circ-ZKSCAN1 gene in a patient with hepatocarcinoma is detected to find that the expression level of the circ-ZKSCAN1 gene is significantly reduced; hepatoma cells transfected with circ-ZKSCAN1 gene shRNA sequence are compared with control hepatoma cells transfected with an empty vector to find that the proliferation rate, cell migration ratio and cell invasion ability and the like are significantly improved. The circ-ZKSCAN1 gene and a circ-ZKSCAN1 gene expression product are used as a hepatoma diagnostic marker, so that hepatoma diagnosis is more accurate and rapid, and the circ-ZKSCAN1 gene as a target gene for preparing hepatoma treatment drugs provides a new therapeutic target and therapeutic approach.

The invention relates to a biological cellulose skin repair material and a preparation method thereof. The preparation method is characterized in that a bacterial solution is added in a culture emulsion, is uniformly dispersed and is then subjected to static culture to obtain the biological cellulose skin repair material, wherein the bacterial solution is a system by which a bacterial strain is dispersed in water; the culture emulsion is an oil-in-water emulsion; an aqueous phase is a bacterial strain culture solution; the bacterial strain means a microorganism capable of biologically synthesizing cellulose nano fibers. The biological cellulose skin repair material and the preparation method thereof provided by the invention have the benefits that the preparation method is simple and easyto implement, convenient to operate and low in cost; the self structure of the material in the preparation process is easily regulated and controlled; the biological cellulose skin repair material prepared finally has the advantages of being good in biocompatibility, excellent in mechanical property and optimal in water-absorbing capacity and water holding capacity; the moisture volatilization canbe reduced, a wet state is kept, external bacteria is effectively blocked, the adhesion and the proliferation rate of cells are promoted, the cells are guided into the interior of the material, and the vascularization of surrounding tissues is accelerated; therefore, the skin wound repair is significantly accelerated.

The invention relates to a traditional Chinese medicine extract capable of obviously improving CIK (Cytokine-Induced Killer) cell proliferation rate. The extract is prepared from the raw materials of red ginseng, radix ophiopogonis and milk vetch according to a certain weight ratio. A formula of the extract comprises the steps as follows: the red ginseng, the radix ophiopogonis and the milk vetchare added with water and decocted, and are subjected to ethanol precipitation and filtered; ethanol is recovered; the extract liquid is enriched with macroporous resin columns, eluted to be colorlesswith distilled water and eluted with ethanol; the eluant is collected; the extract liquid is filtered with a microporous filtration film; and the filter liquor is condensed, the pH value is adjusted,the filter liquor is filtered again with the microporous filtration film, sterilized, and subpackaged, and then the traditional Chinese medicine extract is obtained. Pharmacological experiments show that the traditional Chinese medicine extract can remarkably improve the proliferation rate of CIK cells.

The invention discloses a method for rapidly breeding fritillaria pallidiflora by using a tissue culture technique. The method mainly comprises the steps of selecting and disinfecting explants, treating materials, inducing callus, carrying out subculture, carrying out rooting culture, seedling hardening, transplanting and the like, wherein the formation rate of the callus of the fritillaria pallidiflora is 93%, the proliferation rate is 6 times, the rooting percentage is 92%, the final survival rate is 90%, and the quality and the output of the fritillaria pallidiflora are both greatly improved; and moreover, the method simulates the growing environment of the fritillaria pallidiflora, the temperature is low in the culture process, chernozem is mainly taken as the culture medium, and the quality is greatly improved as the fritillaria pallidiflora grows in such a simulated wild environment.

The present invention relates to a human mesenchymal stem cell serum-free medium, the main components of the human mesenchymal stem cell serum-free medium include a basal medium and an additive; the mesenchymal stem cell serum-free medium has a clear composition and no animal-derived component. A mesenchymal stem cell product produced by the human mesenchymal stem cell serum-free medium is used clinically without causing anaphylactic reaction; and the serum-free medium is simple in preparation method, does not require any special equipment, and is easy to be mass-produced; and mesenchymal stemcells produced by the human mesenchymal stem cell serum-free medium have good cell homogeneity and high proliferation rate. After repeated passages, detection results of cell surface markers are up to standard, and the cells still have good capability of differentiation into osteoblasts, adipocytes and chondrocytes.

Provided is yarn for a cell culture scaffold. The yarn includes ply-twisted fiber strands, and to prevent density-dependent inhibition of cultured cells and increase a cell-contacting specific surface area, at least a part of the plurality of twisted fiber strands are untwisted such that an open space is formed between the fibers. A cell proliferation rate and cell viability may be increased by creating microenvironments suitable for migration, proliferation and differentiation of the cultured cells using the yarn. A large quantity of cells may be simultaneously cultured by creating a cell proliferation space as large as possible in a scaffold space having a limited cell proliferation space, and cell proliferation may be steadily maintained by preventing the inhibition of cell proliferation due to intercellular contact. The cells cultured may be cultured to have a shape / structure suitable for application to an in vitro experiment model or implantation into an animal body.

The invention relates to a serum-free medium used for culturing Vero cells. The serum-free medium is composed of, by volume, 85 to 95% of a base culture solution, 2 to 5% of an amino acid solution, 2 to 7% of a serum alternative factor solution, 0.1 to 0.5% of an antioxidant solution, 0.8 to 2% of a yeast extract solution, and 0.1 to 0.5% of an ethanol amine solution. The invention also provides a preparation method of the serum-free medium. In the preparation method, the base culture solution, the amino acid solution, the serum alternative factor solution, the antioxidant solution, the yeast extract solution, and the ethanol amine solution are mixed at the above ratio so as to obtain the serum-free medium. The serum-free medium used for culturing Vero cells is capable of promoting rapid attachment of Vero cells; cell morphology of the Vero cells cultured with the serum-free medium can be maintainer preferably, and cell proliferation rate is higher; the composition of the serum-free medium is simple; cost is reduced; and preparation is convenient.

The invention belongs to the field of plant biotechnology, relates to medicinal plant cultivation technology and particularly relates to a method for culturing dendrobium candidum protocorms. The method comprises the following production steps in sequence: preparation, protocorm germination and proliferation, transitional observation and culture, and bioreactor production. The method for producing the dendrobium candidum protocorms by using an intermittent immersion bioreactor is carried out under sterile conditions, the sterilization treatment method is damp and hot sterilization at high temperature and high pressure, in the technical scheme, the intermittent immersion bioreactor is used for culturing the dendrobium candidum protocorms, so that a high proliferation rate is obtained within a short time, the culture period of the dendrobium candidum protocorms is shortened and the quality of the cultured dendrobium candidum protocorms is high; the method is simple to operate and is automatic, thereby saving time and labor; meanwhile, the use of culture dishes is reduced to save the manpower cost and the culture cost and achieve a low-cost culture method, and thus the method is suitable for industrial promotion.

The invention discloses a tissue culture and regenerated plant in vitro mycorrhization method for pinus massoniana, which belongs to the technical field of biotechnology and modern forestry. The method comprises the following steps of: taking a terminal bud of an aseptic seedling of the pinus massoniana as an explant, inducing to produce a large quantity of multiple shoots with strong growth first, further inducing the generation of an adventitious root, transferring a plant with the macroscopic adventitious root onto a perlite substrate under the condition of asepsis after the induction of the adventitious root is finished, and inoculating ectomycorrhizal rungi Pisolithustinctorius at the same time to realize the mycorrhization in a bottle. The mycorrhization improves the quality of a root system of the adventitious root of a tissue culture regenerated plant of the pinus massoniana, and ensures that the average lateral root number and the average root length are greatly improved. The mycorrhization also makes the state of the adventitious root greatly changed, and a fungal mantle obtained by closely arranging more than ten layers of mycelia is formed on the surface of the adventitious root. Through the method, the induction rate of the multiple shoots of the pinus massoniana reaches 95 percent, the proliferation rate reaches 430 percent, the rooting rate reaches 85 percent, and the transplantation survival rate is improved to more than 85 percent from 65 percent.

The invention aims to provide a bacillus licheniformis and application thereof. The preservation number of a bacterial strain of the bacillus (Bacillus lincheniformis) LV005 is CCTCC M 2013262. The bacillus licheniformis LV005 disclosed by the invention is screened from an alimentary canal of a prawn, and has the advantages of improving the composition and the number of dominant microbial populations in the alimentary canal of the prawn, forming and maintaining good microecological balance of the alimentary canal, and resisting the adhesion and the proliferation of viruses in the alimentary canal through nutrition competition, space competition, bacteriocin secretion and the like. The bacterial strain can activate the expression of prophenoloxidase, superoxide dismutase, lysozyme, heat shock protein, lipopolysaccharide-glucan conjugated protein, signal transduction, transcriptional activation factors and the like of the prawn with relevant pathogenic infection resistant molecules, and retard the proliferation rate of the viruses in the body of the prawn, so that the bacterial strain has various biological functions of improving the activity of the protease, the lipase and the amylase of the prawn, promoting the digestion capacity of the prawn and the digestion for feed nutrients, improving the stress response resistant capacity of the prawn and the like.

The invention belongs to the technical field of plant seedling propagation and discloses a micropropagation method of tilia miqueliana. The method comprises steps as follows: A, culture of seedling of the tilia miqueliana; B, adventitious bud induction; C, high-voltage electrostatic field processing; D, subculture and rooting; E, acclimation and seedling hardening. A large quantity of tilia miqueliana explants easy to disinfect can be obtained, the success rate of primary culture is higher than 98%, the proliferation rate reaches 113 times, the survival rate reaches 95%, the problem of shortage of the tilia miqueliana in the market is effectively solved, and the application prospect is wide.