70results about How to "Improve sticking rate" patented technology

The invention discloses a polylactic acid self-curled and self-bonded composite cigarette tow and a preparation method thereof. The preparation method comprises the following steps: drying polylacticresin; adding a spinning assistant for the polylactic resin into conventional polylactic resin, adding the spinning assistant to polylactic resin containing a plasticizer, and performing melting plasticization to obtain a polylactic resin melt; spraying the melt from a spinneret hole to form single composite fibers; cooling a polylactic acid two-component cigarette tow; oiling the two-component tow, and carrying out drafting and tension heat setting to obtain the polylactic acid self-curled and self-bonded composite cigarette tow. The single fibers in the tow is one of a side-by-side, orange-slice, sheath-core and sea-island composite structures, one of the components of the side-by-side type or the orange-slice type structure is the polylactic resin containing the plasticizer, and the remaining component is the conventional polylactic resin and a composition thereof. The preparation method has the advantages of simplification of the production process, avoiding of mechanical and thermal damages, increase of the rod yield of the polylactic fiber tow, and reduction of the production cost; and the polylactic acid self-curled and self-bonded composite cigarette tow has a good biodegradability.

The invention relates to a polylactic acid filter stick paper element and a manufacturing method thereof, in particular relates to a biodegradable polylactic acid filter stick paper element and a preparation method thereof, and belongs to the technical fields of tobacco tar filter elements and manufacturing methods. The polylactic acid filter stick paper element is prepared from the following raw materials in parts by weight by virtue of melt spinning: 100 parts of polylactic acid and 5-15 parts of a bonding agent, wherein a crystallization nucleating agent accounts for 0.15-0.5% of the mass percentage of polylactic acid; the ratio of a laevo isomer in the polylactic acid is 95-99%; the crystallization nucleating agent is a mixture of a substituted aryl lithium phosphate salt Na-03 and nano calcium carbonate, the substituted aryl lithium phosphate salt Na-03 accounts for 0.10-0.30% of the mass of the polylactic acid, and the nano calcium carbonate accounts for 0.05-0.2% of the mass of the polylactic acid. The materials used by the paper element are rich in resource, the manufacturing cost is low, the curling property index, tensile strength, filtration efficiency and quality characteristic of the filter stick paper element prepared by the method are close to those of cellulose acetate smoke filter material, and the polylactic acid filter stick paper element is biodegradable.

The invention discloses a preparation method of a complex live vaccine for porcine reproductive and respiratory syndrome, belonging to the technical field of veterinary biological products. The preparation method comprises the following steps: (1) culturing master cells; (2) performing suspension culture on vaccine preparation cells; (3) culturing and harvesting viruses; (4) performing mixed adsorption on the harvested full-suspension virus solution and chicken anti-porcine reproductive and respiratory syndrome virus egg yolk antibody; (5) adding the adsorbed antigen-antibody compound into an immunoenhancer-containing freeze-drying protecting agent, sub-packaging and then performing freeze vacuum drying to prepare the complex live vaccine for the porcine reproductive and respiratory syndrome. The method is used for preparing the vaccine, and has the characteristics that the production period can be shortened, the personnel allocation is reduced, the pollution probability is low, the floor area is small, the virus titer is high, the difference between the prepared vaccine batches is small, the product has stable and high-efficiency quality, the side effect is small and the quality of the vaccine can be completely improved.

The invention discloses a method for culturing human airway epithelial cells. The method comprises the following steps of: (1) acquiring human airway epithelial cells which are subjected to primary culture; and (2) acquiring human airway epithelial cells which are subjected to subculture, namely cleaning by using a phosphoric acid buffer solution when the fusion density of primary cells reaches 70 to 90 percent, adding a 0.25 percent trypsin solution, digesting at room temperature for 5 to 10 minutes, collecting cell suspension when cells are retracted and suspended, adding an equal amount of solution containing a protease inhibitor, performing centrifugal collection on the cells, inoculating in a novel culture dish containing a complete medium at the concentration of between 1 and 6*10<6> cells / ml, culturing, and thus obtaining the human airway epithelial cells which are subjected to subculture when the cells grow to a logarithmic growth phase after 3 to 5 days. The method for culturing the human airway epithelial cells has a stable technology and is high in repeatability; and the cultured cells have high purity and uniform morphology, grow well and can be subjected to continuous passage.

The invention discloses human mesenchymal stem cell preservation transportation liquid which is prepared from 1 to 15 mg / ml of sodium chloride, 1 to 15 mg / ml of sodium gluconate, 1 to 10 mg / ml of sodium acetate, 0.1 to 9.9 v / v% of human AB serum, 0.1 to 25 mg / ml of ginsenoside Rb1, 1 to 100 ng / ml of transferring, 1 to 100 ng / ml of reduced glutathione and 0.01 to 1 mmol / l of glutamine. The invention further discloses application of the liquid in preservation and transportation of human mesenchymal stem cells. The human mesenchymal stem cell preservation transportation liquid can reduce the clustering phenomenon of the human mesenchymal stem cells in a transportation process and keep high attachment rate and activity, so that the human mesenchymal stem cell preservation transportation liquid can be conveyed to all parts of the country and surrounding countries through modern vehicles such as an airplane, and the clinical use range of the human mesenchymal stem cells is greatly enlarged.

The invention discloses a tow with a high crimp index for a polylactic acid cigarette and a preparation method for the tow. The tow is prepared from the following raw materials by a melt spinning method: 99-100 percent by weight of polylactic acid, 0.1-0.5 percent by weight of nanoscale titanium dioxide, 0.1-0.5 percent by weight of hydrated magnesium silicate ultrafine powder, wherein the linear density of the tow is 3.5-5.0 ktex, the linear density of a single filament is 2-5 dtex, the crimp number is 18-28 / 25 mm and the crimp index is 0.45-0.65. According to the tow and the preparation method disclosed by the invention, a nucleating agent is added into a polylactic acid raw material, so that the crystallization rate is increased and the crystallinity degree and the thermal performance of fibers are improved, and further the crimp temperature in the subsequent spinning crimp work procedures is increased to 85-95DEG C and the crimp performance is improved; and the crimp index of the whole tow is improved to about 0.55. The tow with high crimp index for the polylactic acid cigarette has better crimp performance; and under the same conditions, unit weight tows can be used for preparing more filter rods, and thus the economic benefits are increased.

The invention discloses a process for producing a filter rod by utilizing a KDF4 forming unit. The process comprises the following steps: throwing raw tows into an AF4 tow opening device; after spraying a plasticizer, delivering the tows to the KDF4 forming unit by a delivering roller to produce the filter rod; after the filter rod is formed, detecting the filter rod, putting the filter rod into a disk and storing the filter rod; and then delivering the filter rod to a cigarette making machine. The process has the following beneficial effects: the process parameters are optimized; aiming at the tows of different specifications (such as 3.5Y34000, 3.0Y35000, 3.9Y31000, 3.0Y32000, 2.4Y34000 and the like), the optimal process parameter is screened out through experiments; the stability of indexes such as resistance to suction of the KDF4 forming unit and the yield of the filter rod produced by utilizing the KDF4 forming unit are improved on the original bases; and the purpose of reducingthe processing consumption is achieved.

The invention discloses a primary rat or mouse gastric mucosal epithelial cell isolation and culture method. A perfusion method is innovatively introduced into the primary rat or mouse gastric mucosal epithelial cell isolation and culture method, and sufficient perfusion without dead corners is achieved. Prepared compound enzyme digestive juice avoids the problems of low cell viability and poor yield and is capable of digesting more evenly and thoroughly as compared with preheated digestion of traditional single collagenase. Within 12 hours of cell inoculation adherence, a cell culture medium with a pH value of 6-6.5 is adopted, cell adherence rate can be increased greatly and adherence effects are better. The primary rat or mouse gastric mucosal epithelial cell isolation and culture method has the advantages of large total cell number, high isolation degree, high living rate and low contamination probability by bacteria, fungi and other cells as well as capability of achieving subculture so as to provide a favorable cell culture scheme for related fundamental researches.

The invention discloses a hollow polylactic acid fiber for a cigarette cooling filter section and a preparation method of the hollow polylactic acid fiber. The method comprises the follow steps: withfood fiber-grade polylactic acid as a raw material, drying, melting, spinning, cooling, oiling, drafting, crimping, drying and shaping are performed, and the fiber is prepared. In the smoking processof a cigarette with a filter tip prepared from the hollow polylactic acid fiber, the hollow polylactic acid fiber has high adsorption and absorption capability to polar and non-polar smoke component gases; smoke is heated after passing through the filter tip, the polylactic acid material absorbs heat during phase transition, and the prepared hollow fiber has large diameter and large smoke passage,the contact exchange specific surface of the cigarette filter tip and the smoke is increased, the smoke temperature is reduced, and the harm reduction effect is achieved.

The invention discloses a separation and culture method of bovine umbilical cord mesenchymal stem cells. The method comprises the following steps: taking fetal bovine umbilical cord and separating cells by a tissue block adherence method; after the cells are separated, cultivating the cells in a complete culture medium; when cell fusion degree reaches 80%-90%, digesting for 1-3 minutes, terminating the digestion and then blowing and beating to obtain the bovine umbilical cord mesenchymal stem cells, wherein the complete culture medium is a serum-free culture medium added with EGF and PDGF, concentration of the EGF is 5-10ng / mL and concentration of the PDGF is 5-10ng / mL. According to the invention, the tissue block adherence method is utilized to perform a primary separation to the bovine umbilical cord MSC and EGF and PDGF are added into the serum-free culture medium to perform culture amplification, so as to successfully obtain bovine umbilical cord MSC with self renewal and differentiation potentials. Addition of EGF and PDGF in the culture system not only promotes in-vitro amplifications of the cells, but also inhibits differentiation of the cells in in-vitro cultivation.

The invention discloses a method for quickly separating and culturing fibroblast from human skin tissues. The method comprises the following steps of cleaning and shearing fresh foreskin tissues afteroperation of adults or children, and sequentially adopting solutions of dispase II and dispase I to digest under the condition of oscillation in a water bath at the temperature of 37 DEG C; and collecting skin tissue blocks through centrifuging; enabling a culture medium to resuspend, and culturing in a culture box with CO2 (carbon dioxide) volume concentration of 5% at the temperature of 37 DEGC. The method has the advantages that the temperature of the digestion function of the dispase I and the dispase II is increased from 4 DEG C to 37 DEG C, the activities of the dispase I and the dispase II are respectively improved, the contact areas between the dispase I as well as the dispase II and a corresponding primer are increased by the oscillation in the water bath, the enzymatic reaction is favorably performed, the digestion time of the dispase I as well as the dispase II is greatly shortened, and the separating and culture speed of the fibroblast is improved; the damage of enzyme to tissue cells is decreased, the wall attaching rate of the cells is improved, and the primary fibroblast can complete passage after culturing for 6 to 8 days.

The invention relates to a culture medium for ricefield eel germline stem cells. The culture medium comprises a basic culture medium and FBS, wherein the concentration of the FBS is 5-8%. The invention also relates to a method for separating and culturing the ricefield eel germline stem cells. The method comprises the step of culturing the ricefield eel germline stem cells by using the culture medium. According to the culture medium disclosed by the invention, the adherence efficiency of the ricefield eel germline stem cells can be improved, differentiation of the ricefield eel germline stem cells is prevented, and the proliferation function of the ricefield eel germline stem cells is not influenced. By the method disclosed by the invention, ricefield eel female germline stem cells and male germline stem cells which have higher purity can be separated and obtained in five days by using gonad tissues of ricefield eels. The obtained germline stem cells have typical stem cell morphology,typical stem cell clone can be formed, alkaline phosphatase staining is positive, and related molecular markers of the stem cells are also highly expressed. After the germline stem cells obtained by the method disclosed by the invention are cryopreserved, the germline stem cells can be successfully recovered.

The invention discloses a composition containing a recombinant human fibronectin peptide. The composition is composed of an active substance and an excipient, the active substance includes fibronectin, lysozyme, arginine / lysine polypeptide and tripeptide-1 copper. The weight ratio of the fibronectin to the lysozyme to the arginine / lysine polypeptide to the tripeptide-1 copper is (0.01-10) to (0.01-10) to (0.01-10) to (0.01-10). The composition has remarkable efficacy in promoting wound healing and smoothness, controlling oil and water and the like. The composition can be further prepared intosemi-solid preparations such as lyophilized powder, nano microspheres, nano liposomes, aqueous agents, gels after being compounded with appropriate excipients, and is used as an active additive in thefield of tissue engineering, pharmacy and beauty skin care.

The invention relates to the technical field of cell isolated culture, particularly a digestive enzyme composition and application thereof, and an isolated culture method of umbilical epithelial cells. The digestive enzyme composition comprises hyaluronidase, collagenase VIII and pancreatin. The umbilical epithelial cell primary isolation adopts the hyaluronidase, collagenase VIII and pancreatin for combined digestion. The digestive enzyme composition has the advantages of mild properties, low cell trauma and high enzymolysis rate and greatly enhances the epithelial cell isolation efficiency.

The invention relates to the medical field, particularly to a composition and its use, an umbilical cord preservation preparation and a preparation method thereof. According to the invention, physiological saline is taken as the basic ingredient of umbilical cord preservation liquid, umbilical cord blood plasma is used as the nutritional ingredient source, as a natural substance, umbilical cord blood plasma not only provides multiple nutritional ingredients, but also can maximumly maintain the umbilical cord vitality in order to maintain a microenvironment similar to that in vivo. Lactobionic acid, hydroxyethyl starch and other macromolecular substances are added to avoid swelling death of the umbilical cord in the preservation process. The preservation liquid can be used for transportation and preservation of the preparation at low temperature. Thus, adequate nutrients are supplied to in vitro umbilical cord, and also the seed cell vitality, the original cell morphology and biological characteristics are well maintained.

The invention provides a mesenchymal stem cell resuscitation protective solution, a preparation method and resuscitation method. The resuscitation protective solution is prepared from a cell culture stock solution, a fresh culture medium, L-carnitine and mycose. The resuscitation method comprises the following steps: taking out a cryopreserved mesenchymal stem cell, placing at the temperature of 42-45 DEG C until the mesenchymal stem cell starts to thaw; adding the resuscitation protective solution, uniformly mixing, centrifuging, and washing precipitate; adding the resuscitation protective solution into the washed mesenchymal stem cell, regulating the cell density, culturing, replacing a complete culture medium after 24 hours, and continuing culture by a conventional technology. Accordingto the resuscitation method, the resuscitation protective solution consisting of specific ingredients can be used for resuscitating cells and used as a cell washing solution due to the synergistic effect of the ingredients. The mesenchymal stem cell is resuscitated by combining the resuscitation protective solution and the resuscitation method, the cell reclamation rate and biological activity after cell resuscitation can be remarkably improved.

The invention discloses a primary isolation method of animal skin fibroblasts. Disinfected animal skin is selected, and a full-layer skin wound is formed; an injured skin area is taken down after the skin is injured within 3-5 days, subcutaneous tissue and extra blood vessels are removed completely, and the skin is cleaned with PBS (phosphate buffer saline) containing 2% of a penicillin-streptomycin solution for 2-3 times; the skin is cut into tissue pieces ranging from 0.1 mm<3> to 0.5 mm<3> in size; PBS containing 1% of the penicillin-streptomycin solution is poured, centrifugation is performed at the low speed of 1,000 rpm / min for 5 min, and residual floating tissue pieces and a residual supernatant are removed; a cell culture dish covered with 0.2% of gelatin is evenly inoculated with centrifuged tissue pieces, a cell culture fluid containing 2 ml of a DMEM (Dulbecco modified eagle medium), 10% of the FBS and 2% of the penicillin-streptomycin solution is added, and the culture dish is placed in a 5% CO2 incubator at the temperature of 37 DEG C for culture. A skin injury healing mechanism is used, a proper healing period is selected, a large number of proliferative fibroblasts are obtained, meanwhile, removal of the subcutaneous tissue facilitates climbing out of the fibroblasts of corium layers, the number of cells can be increased, the cells can climb out more easily, the cell climbing-out cycle is shortened, and the adherence efficiency is enhanced.

The invention provides a preparation method of placenta decidua mesenchymal stem cells. The preparation method comprises the following steps: s1, putting a placenta decidua material into a sterile tray containing 75% alcohol; s2, cleaning the placenta decidua material with normal saline to remove alcohol; s3, conducting cleaning through a D-Hanks solution, removing blood vessels on each small block and squeezing out blood in the blood vessels; s4, cutting the placenta decidua material into small pieces by using scissors, cleaning the small pieces by using normal saline, and then cutting the small pieces into tissue blocks; s5, performing digesting by using a combined digestive enzyme; s6, diluting the tissue mixture by using PBS (Phosphate Buffer Solution) with the same volume, and takingsupernate suspension; s7, centrifuging the supernate suspension to obtain a precipitate, resuspending the precipitate with an erythrocyte lysis solution, performing standing for lysis, performing centrifuging to obtain a precipitate, resuspending the precipitate with a culture solution, and performing inoculating to a culture dish; and s8, carrying out serum-free amplification passage on the precipitate in the culture dish to obtain passage clinical-grade placenta mesenchymal stem cells. The whole culture period is short, the efficiency is high and the stability is good.

The invention discloses an in-vitro separation and cultivation method of deer dermal papilla cells and application thereof, and belongs to the technical field of cytobiology. The in-vitro separation and cultivation method comprises the steps that the complete hair follicle in deer skin is separated under an integrated microscope; then collagenase is used for digesting the hair follicle; a hair bulb containing a papilla is cut off; a hair root sheath is stripped to obtain the papilla; the papilla is collected; then the collagenase is used for digesting a basilar membrane; and the papilla cells are acquired after development. The in-vitro separation and cultivation method has the advantages that the cultivation success rate is very high, and the adherence rate of the papilla and the cell yield are obviously improved. By the adoption of the cultivation method, about 100 pure dermal papillae can be obtained within about four hours, and a larger number of dermal papilla cells can be obtained when the dermal papillae are subjected to inoculated culture for about seven days. The in-vitro separation and cultivation method is suitable for in-vitro separation and cultivation of the deer dermal papilla cells.

The invention discloses an efficient pre-treatment device for culturing human amniotic mesenchymal stem cells and a culture method thereof. The efficient pre-treatment device comprises an upper cover, a shell, a cutting assembly, a digestion assembly and a cleaning centrifugal part, wherein the shell is hinged to the upper cover; the cutting assembly located at an opening of the shell is matched with the upper cover; the digestion assembly is located in the middle of the shell; and the cleaning centrifugal part is located on the lower portion of the shell and comprises a U-shaped cabin body, a stirring paddle and a micromotor. The invention also provides a cell pre-treatment and culture method based on the efficient pre-treatment device. The method comprises the following six steps: human amniotic membrane cutting and separating; enzymic digestion; centrifugal purification; culture medium preparation; two-dimensional culture; and subculture. The cell culture method based on the efficient pre-treatment device disclosed by the invention is stable in technology and good in repeatability, avoids the problems of low cell activity and poor yield, improves the cell activity and yield greatly, simplifies the operating process, and is flexible and convenient to use.

The invention discloses an efficient pre-treatment device for culturing human umbilical cord mesenchymal stem cells and a related application thereof. The efficient pre-treatment device comprises a turnover upper cover, a cutting assembly, a digestion assembly and a cleaning centrifugal part, wherein a cutter groove is formed in the upper cover; the cutting assembly located at an opening of a shell is composed of latticed blades and an elastic seal ring and the latticed blades and the cutter groove are correspondingly arranged; the digestion assembly is located in the middle of the shell; and the cleaning centrifugal part is located on the lower portion of the shell. The invention also provides a cell pre-treatment and culture method based on the efficient pre-treatment device. The method comprises the following six steps: human umbilical cord cutting and separating; enzymic digestion; centrifugal purification; culture medium preparation; two-dimensional culture; and subculture. The cell culture method based on the efficient pre-treatment device disclosed by the invention is stable in technology and good in repeatability, avoids the problems of low cell activity and poor yield, improves the cell activity and yield greatly, simplifies the operating process, and is flexible and convenient to use.

The invention discloses a high-flux cell culture apparatus and a manufacturing method thereof. The high-flux cell culture apparatus comprises a cell culture room which is formed by a plurality of independent cell culture layers which are isolated from one another by clapboards; each cell culture layer is internally provided with a culture medium feed and discharge hole and a sealing plug; a ventilation port provided with a filter is inserted into the rear of the sealing plug; the culture medium feed and discharge hole is connected with a feeding device by a pipeline; the feeding device is respectively connected with a culture solution storage tank and a cell collection bottle by the pipeline; and a feed power device is a single-tube or multi-tube peristaltic pump which is used for conveniently adjusting the flow velocity and the flow rate. The high-flux cell culture apparatus can be used for confirming the cell culture dose according to the need and can conveniently switch different culture mediums, thus meeting the demands of different cell culture stages and conveniently adjusting the cell culture dose; and the high-flux cell culture apparatus can be used for research and development as well as actual production, and has wide application scope.

The invention discloses a preparation method of adipose-derived stem cell multi-cell active factor freeze-dried powder. The preparation method comprises the following steps: S1, collecting adipose tissue of a human body; S2, conducting primary separation, culture and passage of adipose-derived stem cells; S3, carrying out large-scale amplification on mesenchymal stem cells and collecting supernate; S4, preparing a concentrated stock solution of stem cell active factors; and S5, preparation of freeze-dried powder. The multi-cell active factor freeze-dried powder prepared by the preparation method disclosed by the invention includes a large number of cellular active factors, and is safe and reliable to use, free of any adverse reaction, high in yield, good in activity, easy to store and capable of remarkably promoting healing of deep cell tissues of damaged skin and fading scars and acne marks.

The invention provides a preparation method for PEEK with a topological pattern on the surface. The preparation method comprises the following steps of: (1) placing a PEEK material on a template witha topological pattern on the surface, carrying out compression molding on the PEEK material through cold pressing, hot pressing or a method of firstly carrying out cold pressing and then carrying outheating, and meanwhile, transferring the topological pattern on the template to the surface of the PEEK subjected to compression molding; and (2) separating the template from the PEEK subjected to compression molding to obtain the PEEK with the topological pattern on the surface. According to the method provided by the invention, the autonomously designed topological pattern can be formed on the surface of the PEEK in a large area, the formed topological pattern has relatively high resolution, and the elasticity modulus of the patterned PEEK can be adjusted by adjusting the extrusion forming condition so as to adapt to different application requirements.

The invention discloses a saccharomyces cerevisiae expressed long-acting recombinant fibronectin and an application of the long-acting recombinant fibronectin in cosmetics. The nucleotide sequence of the long-acting recombinant fibronectin is shown as Seq ID NO.1, and the long-acting recombinant fibronectin comprises a functional fragment of FN and HAS; the functional fragment of the FN comprises an FNIII1C unit; and the FNIII1C unit is a functional fragment at the C end of the first III-type FN repetitive unit in the FN. FNIII1C is a functional fragment at the C end of a first III type FN repetitive unit in natural human FN, and has the functions of resisting tumors, resisting metastasis and inhibiting angiogenesis. The protein can also enable FN in a living body to be subjected to super-polymerization, and the cell adhesion promoting and cell migration resisting capabilities of the FN are greatly enhanced. The fusion HAS effectively improves the stability of the recombinant protein. The long-acting recombinant fibronectin disclosed by the invention can be used for improving the adherence rate and convergence rate of cells, shortening the cell convergence time, enabling the morphological structure of the cells to be good, enhancing the metabolic rate and remarkably improving the protein synthesis speed, and has the effect of promoting cell repair.

The invention provides a separation and culture method of renal podocyte of a mouse. The separation and culture method comprises the following steps of (a), intraperitoneally injecting 2% pentobarbital sodium to execute the mouse, sterilizing the mouse with alcohol, taking out kidney tissue of the mouse, putting the kidney tissue into a precooled sterile PBS (Phosphate Buffer solution) containing double antibodies, soaking and washing the kidney tissue, and peeling and removing a renal capsule; (b), mechanically dissociating the kidney tissue, then digesting the kidney tissue by a preheated mixed enzyme solution, and terminating digestion by a mixed complete culture solution; (c), obtaining the renal podocyte by adopting a difference sieving method, inoculating the podocyte in a treated cell bottle, and culturing the podocyte; (d), digesting a cell, which is cultured for 3d, through pancreatin, then filtering the cell once again by a sieve mesh, centrifuging filtrate to remove supernatant fluid, and resuspending and inoculating an obtained mixture in the treated cell culture flask; (e), identifying the renal podocyte by cell immunofluorescence. The separation and culture method of the renal podocyte of the mouse, which is provided by the invention, is good in repeatability, and is simple and convenient to operate; moreover, the high-yield and high-purity renal podocyte of the mouse can be obtained.

The invention discloses a human endometrial stromal cell separation and primary culture method which comprises the following steps of digesting human endometrial tissues by adopting pancreatin withoutcontaining EDTA; during screen filtering of the tissues after digestion, poking filtering tissues to enable the filtering tissues to perform full filtering; and adopting a 1640 culture medium to be used for culture of primary endometrial stromal cells. Compared with the existing method, the method greatly increases the acquisition amount of the primary endometrial stromal cells and the success rate of cell isolated culture, is more efficient and economic and provides a stable and reliable in-vitro cell model to clinical study of endometrial diseases and development of drugs. In addition, an inventor also observes and compares differences between normal endometrial stromal cells and ectopic endometrial stromal cells in the aspects of cellular morphology, the growth characteristic, the biological behavior and the like.

The invention discloses an immortalized urine-source cell strain and a building method thereof and belongs to the technical field of biology. The immortalized urine-source cell strain is obtained by transfecting recombinant plasmids carrying immortalization genes or DNA fragments into urine-source cells. The inventor transfects the SV40LT recombinant plasmids into the urine-source cells to build the immortalized urine-source cell strain. A safe and reliable gene editing method is optimally selected by comparison to obtain the immortalized urine-source cell strain. When the immortalized urine-source cell strain is cultured to the 35th generation, cells are quite young and do not have an evident aging phenomenon; cells are increased logarithmically, and the cell proliferation rate is three times of that of primary-generation urine-source cells which are cultured to the 4th generation. Identification and analysis show that various indexes of the immortalized urine-source cell strain are normal. The cell attachment rate of the immortalized urine-source cell strain recovered after one month of freeze preservation reaches up to 95%, and long-term stable growth and passage can be achieved.

The invention discloses an improved L-15 culture medium and application thereof in culturing prawn muscle cells. The improved L-15 culture medium contains 10 mmol / L of alanine, 3 mmol / L of glycine, 3 mmol / L of serine, 1.5 mmol / L of histidine, 0.5 mmol / L of lysine, 1.5 mmol / L of arginine, 1.5 mmol / L of valine, 1 mmol / L of threonine, 0.5 mmol / L of methionine, 0.5 mmol / L of leucine, 5 mmol / L of aspartic acid, 0.5 mmol / L of glutamine and 1 mmol / L of phenylalanine. The oral liquid also contains taurine, proline, glutamic acid, fetal calf serum and double antibodies, and the pH value of the oral liquid is 7.2-7.4. According to the improved L-15 culture medium, the survival rate and the adherence rate of the primary muscle cells of the penaeus vannamei boone are increased, and the growth state of the cells is improved.