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31 results about "Dmem f12" patented technology
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Method and application for inducing human umbilical cord mesenchyme stem cells to be differentiated into testicular interstitial cells
InactiveCN102174468AHigh differentiation efficiencyGood secretion effectSkeletal/connective tissue cellsViruses/bacteriophagesCorpus luteum graviditatisTesticular Interstitial Cells
The invention discloses a method and application for inducing human umbilical cord mesenchyme stem cells to be differentiated into testicular interstitial cells. The method comprises the following step of culturing human umbilical cord mesenchyme stem cells of patients suffering from adenovirus and carrying mice steroidogenic factor-1 genes in a DMEM-F12 culture solution containing 0.3-3ng / ml of luteinizing hormone, 200-800mu M of dibutyryl cyclic adenosine monophosphate, 5*10<-6>-5*10<-4>M of all-trans retinoic acid (ATRA), 10mU / ml of human chorionic gonadotropin and 2.4uM of adrenocorticotrophic hormone for a week. Induced by the method in the invention, the human umbilical cord mesenchyme stem cells can be differentiated into testicular interstitial cells in vitro and provides important sources of cells for treating testosterone shortage by the cell replacing method or the genetic method.
Owner:JINAN UNIVERSITY
Reprogrammed culture medium and culture method of reprogrammed induced multipotential stem cell
ActiveCN109628383AOvercoming pollutionImprove securityNervous system cellsArtificially induced pluripotent cellsRetinyl acetateInduced pluripotent stem cell
The invention relates to a reprogrammed culture medium and a culture method of a reprogrammed induced multipotential stem cell. The reprogrammed culture medium is composed of a DMEM-F12 basal culturemedium and additive components, wherein the additive components comprise 60-180 microgram / mL of L-ascorbic acid, 5.3-74 micromole / L of hydrocortisone, 3-89 nanogram / mL of sodium selenite, 8-23 micromole / L of Optiferrin, 0.5-7.4 micromole / L of retinyl acetate, 40-60 nanogram / mL plant-derived recombinant human alkaline growth factors, 8-12 microgram / mL of IGF, 0.2-0.6 microgram / mL A-83, 2-6 micromole / L of CHIR99021 and 100-450 micromole / L of sodium butyrate. The use of the reprogrammed culture medium not only can improve the safety of clinical use, but also can improve the efficiency of adult cell reprogramming to induce the multipotential stem cell.
Owner:IREGENE THERAPEUTICS LTD
Mesenchymal stem cell serum-free culture medium
ActiveCN106190964ANormal growthQuality is easy to controlSkeletal/connective tissue cellsOsteoblastCulture mediums
The invention discloses a mesenchymal stem cell serum-free culture medium. The culture medium takes an L-DMEM, DMEM-F12 or alpha-MEM culture medium as a basic culture medium and also includes the following addition components: fibronectin, laminin, transferrin, trypsin, aprotinin, growth hormone, insulin, hydrocortisone, dexamethasone, bFGF, EGF, B27, IGF-I, IGF-II, choline, linoleic acid, sodium selenite, phosphorylethanolamine, glutathione, vitamin C, vitamin E, vitamin B12 and biotin. The culture medium has the advantages of defined composition and controllable quality, and mesenchymal stem cells cultured by the culture medium grow normal; and after multiple passages, the 'dryness' of the mesenchymal stem cells and the ability of the mesenchymal stem cells to be differentiated into osteoblasts, chondrocytes, adipocytes, neurons and other types of cells are still kept.
Owner:GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
Serum-free medium and preparing method and application thereof
InactiveCN105420189AAvoid situations where the growth process is unstableSolve the shortcomings of poor adhesion ability and slow proliferationSkeletal/connective tissue cellsSerum free mediaFiltration membrane
The invention discloses a novel serum-free medium. The serum-free medium comprises, by volume, 0.05-0.2 part of beta-mercaptoethanol, 0.5-2 parts of a non-essential amino acid aqueous solution, 4-6 parts of human mesenchymal stem cell cultural concentrated supernatant and 90-95 parts of a-MEM / DMEM-F12 and recombination human basic fibroblast growth factors with the final concentration of 5-15 ng / ml. A preparing method of the cultural concentrated supernatant includes the following steps that umbilical cord mesenchymal stem cell cultural supernatant is collected; cells, cell debris, impurities and the like are centrifugally removed; filtering is carried out through a micro-filtration membrane; ultrafiltration and concentration are carried out. When cultured through the medium, stem cells still keep the multi-directional differentiation potential and the high multiplication capacity under the long-time culturing condition.
Owner:郭镭 +2
Connexin core sequence-containing amphiphilic polypeptide and application thereof
The invention provides connexin core sequence-containing amphiphilic polypeptide AcN-RADARADARADARADAGGDHLSDNYTLDHDRAIH-CONH2, wherein the amphiphilic polypeptide and the amphiphilic polypeptide with the structure of AcN-RADARADARADARADA-CONH2 are triggered by Ca2+or a cell culture medium DMEM-F12 to automatically form into a compound, i.e. a nanometer-grade fiber material, and the nanometer-grade fiber material is taken as a bracket of the bone mesenchymal stem cells, so that the biological behavior of the cells can be obviously improved, the degenerated intervertebral disc can be repaired, and the original height can be recovered.
Owner:XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
Digestive enzyme liquid for poultry tibial growth plate cartilage primary cell separation and separation culture method of tibial growth plate cartilage primary cells
The invention provides digestive enzyme liquid for poultry tibial growth plate cartilage primary cell separation. A complete medium is used as basic liquid, and is prepared from the following ingredients including collagenase IV with the mass concentration being 80 to 100mg / mL and hyaluronidase with the mass concentration being 80 to 100mg / mL; the complete medium takes a high-glucose DMEM-F12 culture medium as a basic culture medium, and contains the following ingredients including fetal calf serum with the volume concentration being 8 to 12 percent, vitamin C with the mass concentration being 4.5 to 5.5mg / mL and mycillin with the mass concentration being 1 to 10mg / mL. Through the selection on digestive enzyme and the concentration optimization, the cell yield reaches 85 to 95 percent or higher; during the further culture on the digested cells, the cell upward trend is good; meanwhile, the cell viability of the digested primary cell reaches 85 to 95 percent or higher; the cell viability is relatively stable.
Owner:TIBET AGRI & ANIMAL HUSBANDRY COLLEGE
Compositions and methods to culturing neural stem cells with bone marrow stromal cells
The present invention encompasses methods and compositions for enhancing the growth of neural stem cells. Methods for modulating MHC molecule expression on a neural stem cell (NSC) are also included in the invention. Figure 1 is a graph depicting the proliferation of BMSCs in BMSC medium (DMEM-low glucose, 10% lot tested fetal bovine serum) or NSC medium (DMEM-F12, N2-Supplement, EGF 20 ng / ml, bFGF 10 ng / ml, heparin 8 ug / ml, penicillin-streptomycin).
Owner:THERADIGM INC
Cell culture medium for preparing human skin flbroblast sheet and using method of cell culture medium
The invention discloses a cell culture medium for preparing a human skin flbroblast sheet. The cell culture medium comprises the following components: (1) a basic culture medium, namely, H-DMEM (high glucose dulbecco's modified eagle medium) or DMEM-F12; (2) an additive component, namely, vitamin C 50-100 [mu]g / ml, and epidermal growth factors (EGF) 3-20 ng / ml, and (3) serum 5-15%. In the fibroblast basic culture medium, other components are added, so that the cultured fibroblasts form a sheet. The invention further discloses a using method of the cell culture medium.
Owner:陕西九州生物医药科技集团有限公司
Method for isolating mesenchymal stem cells from placental vessel and used digestive enzyme composition
ActiveCN109234229AGood differentiation effectHigh purityCell dissociation methodsSkeletal/connective tissue cellsVascular tissueTissue fluid
The present invention relates to a method for isolating mesenchymal stem cells from placental vessels and used digestive enzyme compositions. The method comprises the following steps: the placenta issterilized with alcohol; Dissecting placental blood vessels from the placenta; Cut into pieces, wash with PBS, filter the residual blood stains to obtain placental vascular tissue; Adding mixed enzymesolution to digest; End digestion, filter and collect tissue fluid; The placental mesenchymal stem cells (P0) were obtained by centrifugation and cultured with DMEM-F12 basal medium was resuspended and the number and viability of nucleated cells were counted. The obtained cells are cryopreserved with cryopreservation protection solution so as to be re-cultured before use, or the passages and / or the mesenchymal stem cells are continued to be subjected to cell identification and / or detection, cryopreservation, library establishment and the like. The method of the invention can effectively improve the efficiency of isolating mesenchymal stem cells from the blood vessels of the placenta.
Owner:BOYALIFE
Cell freezing medium
The invention discloses a cell freezing medium. The cell freezing medium comprises a basal culture medium, glycerin, fetal calf serum and a non-permeable protective agent, and the contents of the components are 80-94 v / v% of the basal culture medium, 5-10 v / v% of glycerol, 1-10 v / v% of the fetal calf serum and 0.01-0.10 w / v% of the non-permeable protective agent, and the basal culture medium comprises RPMI-1640, MEM, high-sugar DMEM, low-sugar DMEM or DMEM-F12; and the non-permeable protective agent comprises sodium alginate, sodium hyaluronate or gamma-polyglutamic acid. Dimethyl sulfoxide isnot contained, a good freezing effect is achieved through different matching of the non-permeable protective agent with glycerin and fetal calf serum, a strong water molecule complexing ability is achieved, the freezing function is similar to the freezing function of dimethyl sulfoxide, but formation of intracellular ice crystals is reduced during freezing; the cell freezing medium has clear added ingredients, high safety, small damage to cells, consistent morphology before and after freezing, a high cell survival rate after cell resuscitation and good proliferative capacity.
Owner:EAST CHINA UNIV OF SCI & TECH
Kit for cell culture
InactiveCN105420186AAvoid situations where the growth process is unstableSolve the shortcomings of poor adhesion ability and slow proliferationSkeletal/connective tissue cellsFiltration membrane3D cell culture
The invention discloses a novel kit for serum-free stem cell culture. The kit comprises a serum-free culture medium, wherein the serum-free culture medium contains 0.05-0.2 part of beta-mercaptoethanol, 0.5-2 parts of a non-essential amino acid aqueous solution, 4-6 parts of human mesenchymal stem cell culture supernatant concentrate, 90-95 parts of a-MEM / DMEM-F12 and human recombinant basic fibroblast growth factors with the final concentration of 5-15 ng / ml by volume. The culture supernatant concentrate is prepared by adopting the following steps of collecting the umbilical cord mesenchymal stem cell culture supernatant, centrifugally removing cells, cellular debris, impurities and the like, performing filtration by using a micro-filtration membrane and performing ultra-filtration concentration. By utilizing the kit for stem cell culture, the multi-directional differentiation potential and stronger proliferation ability of the cells are still kept under the condition of long-term culture.
Owner:郭镭 +2
Serum-free culture medium and preparation method and application therefor
ActiveUS20180362920A1Maintain pluripotencyStrong proliferation capacityCulture processCell culture mediaFiberAdditive ingredient
Provided is a serum-free culture medium, the ingredients of the culture medium comprising 0.05-0.2 parts by volume of β-mercaptoethanol, 0.5-2 parts by volume of non-essential amino acid aqueous solution, 4-6 parts by volume of human mesenchymal stem cell culture supernatant concentrate, and 90-95 parts by volume of a-MEM / DMEM-F12 and recombinant human alkaline fibroblast growth factor of a final concentration of 5-5 ng / ml. The present culture medium is used for carrying out stem cell culture.
Owner:GUO LEI +1
Method for cryopreservation and resuscitation of mesenchymal stem cells
ActiveCN110317781ASuccessful separationSuccessfully and efficiently separateCell dissociation methodsDead animal preservationWater bathsDigestion Treatment
The invention relates to a method for cryopreservation and resuscitation of mesenchymal stem cells. A method for preparation and cryopreservation of the mesenchymal stem cells comprises the followingsteps of disinfection and cleaning, digestion treatment, cell culture, cell passage, addition of a cell cryopreservation solution to the mesenchymal stem cells and cryopreservation in liquid nitrogen.The cell cryopreservation solution contains DMEM-F12, dimethyl sulfoxide, canine blood albumin and the like. The method for cryopreservation and resuscitation of the mesenchymal stem cells comprisesthe following steps that the prepared mesenchymal stem cells and the cell cryopreservation solution are uniformly mixed, then the cell solution is placed in the liquid nitrogen for freezing, and cryopreservation is conducted until the cells are needed; when the cells need to be resuscitated, the cryopreserved cells are taken out of the liquid nitrogen and quickly put into a water bath of 37 DEG Cto make the cryopreservation solution completely thawed, and after the cells are thawed, the cells are immediately transferred to an ice bath to complete the resuscitation of the cells. The inventionfurther relates to the cell cryopreservation solution for the cryopreservation and resuscitation of the mesenchymal stem cells. The method and the cryopreservation solution have excellent technical advantages shown in the description.
Owner:天津瑞博斯生物技术有限公司
Method for improving survival of mesenchymal stem cells after resuscitation and adopted cryopreservation solution
ActiveCN110317782ASuccessful separationSuccessfully and efficiently separateCell dissociation methodsDead animal preservationWater bathsImproved survival
The invention relates to a method for improving the survival of mesenchymal stem cells after resuscitation and an adopted cryopreservation solution. Particularly, the invention relates to the method for cryopreservation and resuscitation of the mesenchymal stem cells. The method comprises the following steps that the prepared mesenchymal stem cells are uniformly mixed with the cell cryopreservation solution, and then the cell solution is placed in liquid nitrogen for freezing until the cells are needed; when the cells need to be resuscitated, the cryopreserved cells are taken out of the liquidnitrogen and quickly put into a water bath of 37 DEG C to make the cryopreservation solution completely thawed, and after the cells are thawed, the cells are immediately transferred to an ice bath of4 DEG C to complete the resuscitation of the cells. The invention further relates to the cell cryopreservation solution for the cryopreservation and resuscitation of the mesenchymal stem cells. The cell cryopreservation solution contains DMEM-F12, dimethyl sulfoxide, canine blood albumin and the like. The invention further relates to a method for preparation of the mesenchymal stem cells from thecanine umbilical cord and cryopreservation of the mesenchymal stem cells. The methods and the cryopreservation solution have excellent technical advantages shown in the description.
Owner:天津瑞博斯生物技术有限公司
Serum-free medium for monkey embryonic stem cell
ActiveCN106754657AMaintain self-renewalMaintain normal stateCulture processCell culture active agentsInsulin activitySerum free media
The invention discloses a serum-free medium for a monkey embryonic stem cell. The serum-free medium comprises a basic medium and additives, wherein the additives are dissolved in the basic medium; the basic medium comprises a DMEM-F12 medium; and the additive comprises L-glutamine, non-essential amino acid, L-ascorbic acid, sodium selenite, beta-mercaptoethanol, human serum albumin, heparin sodium, human transferring, insulin human, a basic fibroblast growth factor, a transforming growth factor beta 1 and an epidermal growth factor. The serum-free medium is capable of maintaining an undifferentiation state and totipotency of the monkey embryonic stem cell within a relatively long time, and is applicable to culture of the monkey embryonic stem cell in both a free-feeding layer system and a feeding layer system.
Owner:北京赛斯达生物技术有限公司
Human induced multipotential stem cells cryopreserved reagent and cryopreserving method thereof
InactiveCN106900695AImprove quality controlSuitable for clinical applicationDead animal preservationHuman albuminDmem f12
The invention discloses a human induced multipotential stem cells cryopreserved reagent and a cryopreserving method thereof. The cryopreserved reagent comprises the following components: 0.03-100 ml of recombinant human albumin, 0.0125 mg / 100 ml of recombinant human epidermal growth protein, 1 ml / 100 ml of beta-mercaptoethanol, 91-100 ml of a DMEM-F12 medium, 8-100 ml of dimethyl sulfoxide, and 5-100 ml of trehalose. The method has the beneficial effects that 1) the component is completely clear and is convenient for quality control; 2) the recombinant human albumin is used for substituting the fetal calf serum of the animal component, can avoid an animal serum-containing component, the cryopreserved reagent is safe and reliable, and is adapted to clinic application; 3) the recombinant human epidermal growth protein, beta-mercaptoethanol, and trehalose in a formula can better protect the cells in a state of dormancy under low temperature environment, ice crystal is not generated in tissue of the cells, cell activity of the cells can be kept, the cell differentiation can be effectively prevented, and the recovery rate of the cryopreserved cell can reach more than 96%; and 4) the cryopreserved reagent belongs to a ready-to-use type, and the usage is convenient.
Owner:北京焕生汇生物技术研究院有限公司
Retinal pigment epithelium epithelial-mesenchymal transition model and application thereof
ActiveCN105112354ATypical pathological process of EMTSimple methodVertebrate cellsArtificial cell constructsSerum freeEssential amino acid
The invention relates to a retinal pigment epithelium epithelial-mesenchymal transition model and application thereof. After being digested with pancreatin, ARPE-19 (acute retinal pigment epitheliitis 19) cells are transferred to a serum-free DMEM-F12 medium containing N2 and non-essential amino acids and are cultured in a Petri dish, and the retinal pigment epithelium epithelial-mesenchymal transition model is acquired. The model is usable for researching a molecular mechanism of retinal pigment epithelium to which epithelial-mesenchymal transition occurs and finding intervened targeting of epithelial-mesenchymal transition.
Owner:TONGJI UNIV
Experimental method for enzymolysis, digestion and acquisition in vitro of rabbit knee cartilage unit
InactiveCN101831402AEasy to getGood repeatabilitySkeletal/connective tissue cellsBiomechanicsKnee Joint
The invention belongs to the technical field of the construction in vitro of tissue engineered cartilage and in particular relates to an experimental method for anzymolysis, digestion and acquisition in vitro of a rabbit knee cartilage unit. The experimental method comprises the following steps of: taking a rabbit knee joint, repeatedly shearing joint cartilages into fragment tissues, washing the fragment tissues, removing supernate and putting the fragment tissues in an aseptic cone bottle for later use; adding dispase enzyme and II type collagenase into the cone bottle in the proportion of 12ml DMEM-F12 culture solution per gram of cartilages, stirring the mixture and digesting to form digesting suspension; and filtering and centrifugating the digesting suspension and adding primary chondrocyte culture solution into the obtained product to prepare aseptic cartilage unit suspension. The experimental method has the advantages that: the acquisition method is simple and the repetitiveness is high; the experimental method more accords with the growing environment and the biological characteristics of the chondrocyte in vitro; and compared with the pure cell, the cartilage unit has obviously improved biomechanics characteristics and the defect of poor biomechanics characteristics of the chondrocyte serving as a seeded cell is overcome.
Owner:THE SECOND HOSPITAL OF SHANXI MEDICAL UNIV +1
Application of exosomes derived from menstrual blood stem cells on preparation of medicament for treating intrauterine adhesion
ActiveCN109893541APromote repairGood treatment effectSurgical drugsMammal material medical ingredientsDiseaseTissue repair
The invention belongs to the field of biological medicines, and particularly relates to application of exosomes derived from menstrual blood stem cells on preparation of a medicament for treating intrauterine adhesion. After primary isolation, culture and passage of the menstrual blood stem cells are carried out, 3rd-6th generations of menstrual blood stem cells are selected for exosomes preparation. When the density of the menstrual blood stem cells reaches 80%, serum components are removed by PBS washing, serum-free DMEM-F12 culture medium is added, and the menstrual blood stem cells are cultured in an incubator at the conditions of 37 DEG C, 5% CO2 and saturated humidity for 24 hours. The supernate is collected, the exosomes are prepared by ultracentrifugation under GMP conditions, andsterile PBS is collected and stored at the temperature of 80 DEG C. The cells have the advantages of being high in survival rate, stable in characters and easy to acquire and expand, and having an obvious treatment effect on intrauterine adhesion. The exosomes can even completely simulate the biological function of the cells to achieve the effects of promoting tissue repair and treating diseases.The exosomes are safer than the cells where the exosomes are derived from, do not lose functions after cryopreservation, can be used for allotransplantation and are suitable for mass production, and have important clinical application values and social and economic benefits.
Owner:SHENGJING HOSPITAL OF CHINA MEDICAL UNIV
Special thawing reagent and thawing method for human induced pluripotent stem cells
ActiveCN107034195AImprove survival rateIncreased versatilityGenetically modified cellsCulture processCuticleDmem f12
The invention discloses a special thawing reagent and a thawing method for human induced pluripotent stem cells. The thawing reagent is free of animal components and comprises 100ng-300ng of recombinant human epidermal growth factors, 1000ng-2000ng of recombinant human blood albumin, 0.1ml-0.25ml of b-mercaptoethanol and the balance DMEM-F12 culture medium, wherein the total volume is 10.0ml. The special thawing reagent and the thawing method for the human induced pluripotent stem cells have advantages that (1) high survival rate (thawing rate) and high activity of the human induced pluripotent stem cells can be guaranteed, and high functional versatility of the cells is guaranteed as well; (2) due to freeness of the animal components, complete exactness and controllability of components can be realized, and high safety in use is achieved; (3) the thawing method is convenient to operate, easy to implement and suitable for wide popularization and utilization.
Owner:焕生汇生物基因技术(北京)有限公司
Isolation and culture method of human adipose-derived stem cells and construction method of stem cell bank
InactiveCN109852583AHigh purityNo differentiationSkeletal/connective tissue cellsLibrary creationPrimary operationCentrifugation
The invention discloses an isolation and culture method of human adipose-derived stem cells. A culture medium is a DMEM-F12 medium added with fetal calf serum with final concentration of 1-20%, bFGF of 1-100 ng / ml and EGF of 1-100 ng / ml. The isolation and culture method includes fat primary operation steps. The fat primary operation steps include steps of crushing, cleaning, digestion, isolation and culturing. The isolation and culture method also includes subculture operation steps. The subculture operation steps include cleaning, digestion, cleaning, centrifugation and culturing. The isolation and culture method of the human adipose-derived stem cells is simple in culture medium composition, short in culture time in the isolation and culture method, fast in growth speed and high in stemcell purity. No stem cell differentiation occurs during the culture.
Owner:CENTURY BIOSTRENGTH BEIJING PTY LTD
Cell culture medium
The invention relates to serum-free cell culture medium, wherein the medium either contains maltose as the sole carbohydrate source, or contains maltose and at least one other saccharide as carbohydrate sources. In a preferred embodiment, the additional saccharide is a monosaccharide, preferably glucose, and the medium comprises DMEM-F12. In a preferred embodiment the cells to be cultured may be CHO or HEK293 cells. Also disclosed are methods of growing and / or culturing a cell using the cell culture medium as described herein, methods of increasing protein yield using the cell culture medium as described herein, and kits thereof.
Owner:AGENCY FOR SCI TECH & RES
Connexin core sequence-containing amphiphilic polypeptide and application thereof
Owner:XIEHE HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI & TECH UNIV
Experimental method for establishing tissue engineering cartilage in vitro by taking knee-joint cartilage unit of rabbit as seed cell
Owner:THE SECOND HOSPITAL OF SHANXI MEDICAL UNIV +1
Method and system for culturing corneal stem cell-like cell line by inducing differentiation of induced pluripotent stem cell using protein ligand
InactiveUS20190177690A1Lower immune responseStrong differentiation abilityCulture processCell culture mediaCulture fluidCell culture media
The present invention relates to a method for culturing corneal stem cell-like cell lines by inducing differentiation of induced pluripotent stem cells (iPSC). The method comprises the following steps:preparing a basal medium comprising DMEM F12, L-ascorbic acid, sodium selenite, and sodium chloride;adding additives for culturing induced pluripotent stem cells to the basal medium;preparing a feeder free culture medium by coating the basal medium with Vitronectin recombinant human protein;adding induced pluripotent stem cells to the feeder free culture medium and culturing them;adding additives for creating an environment for stem cell growth to the feeder free culture medium;adding BMP4 and Wnt3a sequentially to the feeder free culture medium in order to induce differentiation of the induced pluripotent stem cells into corneal stem cell-like cell lines; andculturing the cells differentiated from the induced pluripotent stem cells in PI culture fluid.
Owner:THE CATHOLIC UNIV OF KOREA IND ACADEMIC COOPERATION FOUND
A serum-free chondrocyte culture medium
ActiveCN104630138BEasy to stretchMaintain morphological characteristicsSkeletal/connective tissue cellsInsulin-like growth factorCulture fluid
A serum-free chondrocyte culture medium, in which exogenous additives are added to the basic culture medium. The basal culture fluid is DMEM culture fluid or F12 culture fluid or the mixed culture fluid of DMEM culture fluid and F12 culture fluid, and the exogenous additives are non-essential amino acids, dextran, vitamin C, glutamine, sodium pyruvate, insulin or Insulin-like growth factor, transferrin, growth hormone, triiodothyronine, hydrocortisone, dexamethasone, sodium selenite, beta-mercaptoethanol, lipid concentrate, progesterone, butylenediamine, base fibroblast growth factor, epidermal growth factor, transforming growth factor, bone morphogenic protein, platelet-derived growth factor. The culture medium does not add albumin, which can avoid the potential hidden dangers caused by the use of serum, improve the ability of chondrocytes to secrete extracellular matrix, and is close to the growth and proliferation state in the serum-containing culture medium. The improvement of matrix secretion ability is obviously better than that of serum-containing culture medium.
Owner:SHAANXI RUISHENG BIOTECH
Epithelial-mesenchymal transition model of retinal pigment epithelium and its application
ActiveCN105112354BTypical pathological process of EMTSimple methodArtificial cell constructsVertebrate cellsDmem f12Retinal pigment epithelial cell
The invention relates to a retinal pigment epithelial cell epithelial-mesenchymal transition model and its application. ARPE-19 cells were digested with trypsin, then transferred to serum-free DMEM-F12 medium containing N2 and non-essential amino acids, and cultured in Petri dishes to obtain retinal pigment epithelial cells epithelial-mesenchymal The mesenchymal transition model; this model is used to study the molecular mechanism of epithelial-mesenchymal transition in retinal pigment epithelial cells and to find the intervention target of epithelial-mesenchymal transition. Compared with the prior art, the method of the invention is simple and easy to implement and has good repeatability.
Owner:TONGJI UNIV
Method for high-efficiency amplification of Tembusu virus by chicken hepatocytes
ActiveCN109295010APromote proliferationHigh titerSsRNA viruses positive-senseArtificial cell constructsDmem f12Refrigerated temperature
The invention relates to a method for high-efficiency amplification of Tembusu virus by chicken hepatocytes, wherein the method includes the following steps: (1) cell culture: culturing the chicken hepatocytes in a DMEM-F12 culture medium containing fetal bovine serum with the volume concentration of 10%, after the cells grow on a single layer fully, washing with PBS once, digesting with pancreatic enzymes with the mass concentration of 0.25%, and passaging to a cell culture plate; and (2) high-efficiency amplification of Tembusu virus in the chicken hepatocytes: inoculating the chicken hepatocytes into the cell culture plate and culturing, when the confluence degree is 60% to 70%, inoculating the chicken hepatocytes with the Tembusu virus AHRD2018 strain with MOI of 0.01, culturing in 2 ml of a culture medium containing fetal bovine serum with the volume concentration of 2% for 4 days, collecting 200 [mu]l of a supernatant at a fixed time and cryopreserving in an -80 DEG C refrigerator for standby application, and at the same time, supplementing 200 [mu]l of the culture medium containing fetal bovine serum with the volume concentration of 2% into each hole. The characteristic thatthe chicken hepatocytes can produce high titer virus more prematurely is utilized, and the Tembusu virus is amplified by the chicken hepatocytes having the virus replication rate significantly higherthan that of DF-1 cells and BHK-21 cells.
Owner:YANGZHOU UNIV
A kind of in vitro culture method of rat vascular endothelial cells
InactiveCN104673744BStable and easy to obtainReduced Pollution ChancesArtificial cell constructsVertebrate cellsVascular endotheliumNutrient solution
The invention provides an in-vitro culturing method for vascular endothelial cells of a rat. The method comprises the following steps: a rat aorta is obtained, washed with a sterile PBS (phosphate buffered saline) solution and then soaked in a DMEM (Dulbecco's modified Eagle's medium)-F12 nutrient solution containing heparin; aortic intima is turned out with a sterile needle and thread, and knots are tied at two ends; the overturned aorta is digested in collagenase; digested aorta is placed in a serum-free DMEM-F12 nutrient solution after cultured in an incubator at the temperature of 37 DEG C, and is cut into small aortic slices; the aortic slices are subjected to adherent culture, the aortic intima parts of the aortic slices are soaked in a DMEM-F12 nutrient solution containing heparin and serum, and the nutrient solution is replaced every 48 h; the aortic slices are moved in the third-fifth days, later, the nutrient solution is replaced every day, a large quantity of vascular endothelial cells can be obtained on the ninth day, and the vascular endothelial cells can be used for passage after the fourteenth day.
Owner:武汉市中西医结合医院
Separation and passage method of pregnant rat placenta vascular smooth muscle cell
InactiveCN107058218AReduce the risk of contaminationShorten the timeCell dissociation methodsSkeletal/connective tissue cellsEnzymatic digestionPenicillin
The invention provides a separation and passage method of a pregnant rat placenta vascular smooth muscle cell. The method comprises the steps of taking a pregnant rat placenta, conducting pretreatment, afterwards injecting a mixed solution of a serum-free DMEM F12 culture medium containing agarose, ferroferric oxide and penicillin streptomycin, cutting the placenta into pieces after the mixture is congulated, adsorbing placenta tissue containing iron under the effect of magnetic force, using a collagenase II solution to conduct resuspending, after enzymatic digestion is completed, adding a DMEM F12 total culture medium to passivate collagenase II, and after washing is conducted, adding the DMEM F12 total culture medium again to conduct resuspending, wherein obtained tissue blocks grow sticking to the wall, and then the vascular smooth muscle cell is grown so that subculture can be conducted. According to the separation and passage method of the pregnant rat placenta vascular smooth muscle cell, compared with a traditional method, the placenta vascular smooth muscle cell can be conveniently and efficiently extracted, in the whole process, blood vessels do not need to be dissociated out, the operation is simpler, the time of the tissue for being exposed is shortened, and the risk of cell contamination is lowered.
Owner:CAPITAL UNIVERSITY OF MEDICAL SCIENCES
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