1417 results about "Primary culture" patented technology

Proteolytically stable small molecule β-turn peptidomimetic compounds have been identified as agonists or antagonists of neurotrophin receptors, such as TrkA. A compound of particular interest binds the immunoglobulin-like C2 region of the extracellular domain of TrkA, competes the binding of another TrkA ligand, affords selective trophic protection to TrkA-expressing cell lines and neuronal primary cultures, and induces the differentiation of primary neuronal cultures. The small β-turn peptidomimetic compounds of the invention can activate a tyrosine kinase neurotrophin receptor that normally binds a relatively large protein ligand. Such compounds that bind the extracellular domain of Trk receptors are useful pharmacological agents to address disorders where Trk receptors play a role, by targeting populations selectively.

The invention relates to a method for preparing human umbilical cord mesenchymal stem cells, which comprises the steps of collection, transportation, handover, separation, freezing, recovering, primary culture and subculture of umbilical cords. The method has simple process, ensures low production cost, good freezing effect, little cell damage, low toxic side effect and no harm to human body and reduces the residual of the freezing protection liquid in the recovered cells of tissue blocks; and the prepared mesenchymal stem cells can be applied to the clinical treatment of various diseases.

The invention relates to a method for cultivating autologous umbilical cord mesenchymal stem cells by adopting human umbilical cord blood rich platelet lysate. The conventional cell bank preparing processes adopt blood serum of calves or fetal calves to cultivate, digest, cryopreserve and the like, so that the risk of heterogeneous protein residue certainly exists in the application of clinic umbilical cord stem cells in the future. The method comprises the following steps of umbilical cord blood rich platelet serum preparation, platelet lysate preparation, preparation of serous without platelet, the confirmation of the content of cell factors, namely PDGF-AB, FGF2, TGF-Beta and VFGF in the platelet lysate, the separation and the primary culture of umbilical cord stem cells, the culture and passage of umbilical cord stem cells, the cryopreservation of umbilical cord stem cells, and the karyotype analysis. The method is used for cultivating stem cells.

The present invention demonstrates that VSV-G-pseudotyped lentivirus vectors efficiently transduce AEC in primary culture and in vivo with transduction favored by virus application from the apical side. Transduction efficiency in AEC increased with increasing MOI and greatly exceeded that achieved with a similarly pseudotyped MLV retrovirus vector. The present invention also demonstrates the successful in vivo transfer of genes through lentivirus vector transduction. Mammals injected with lentivirus vector via the trachea expressed the reporter protein in alveolar epithelial cells within 48 to 72 hours after infection.

The invention provides an inducing method for differentiating umbilical cord mesenchymal stem cells (UC-MSCs) into neural stem cells. The inducing method comprises the following steps: carrying out separation and primary culture of UC-MSCs, subculture and amplification of UC-MSCs, induction in vitro of UC-MSC to be transformed into neural stem cells, and differentiation culture of neural stem cells and detection by an immumofluorescence method. The inducing method provided by the invention uses all-transretinoic acid combined with alkaline fibroblast growth factors (bFGF) and epidermal growth factor (EGF) to induce the UC-MSCs to be transformed into neural stem cells, wherein the UC-MSCs still have stronger activity after repeated passage by division growth. The transretinoic acid is combined with cytokines to induce the UC-MSCs to be converted into the neural stem cells, so that the mesenchymal cells are differentiated into non mesenchymal cells cross germinal layer, and the mesenchymal cells probability become more ideal seed cells in the future clinical application.

The invention belongs to the technical field of medicine, and specifically relates to a primary tumor cell culture medium, culture method and application. The primary tumor cell culture method provided by the invention comprises the steps as follows: preparing a primary cell culture medium which comprises the following components: hydrocortisone, EGF, Insulin, a ROCK inhibitor, but not contains cholera toxin, and is an improvement of the existing primary cell culture medium; culturing tumor tissue epithelial cells on laid-out trophoblast cells by using the primary cell culture medium, and enabling the tumor tissue epithelial cells to proliferate rapidly under the combined action of growth factors secreted by the trophoblast cells and nutrient factors contained in the culture medium; and digesting and sub-culturing the tumor tissue epithelial cells when the tumor tissue epithelial cells grow to a cell density of about 80% to 90%. A convenient primary cell culture method is used to obtain immortalized cells possessing the biological characteristics of a patient's own tumor, and the problem of immortalization of the primary culture of tumor cells is solved, thereby realizing personalized treatment for the patient.

The invention relates to a preparation method for a compound fungus agent for specifically degrading an organic matter for a biological enhancing water treatment system. The compound fungus agent is added into a biological treatment process, has high biological activity, a stable effect and low cost, is easy to adsorb on the surface of a carrier and is difficult to lose. The preparation method issimple and practical, and has a short preparation period and low cost. The compound fungus agent comprises the following four strains: 20 to 30 percent of pseudomonas-stutzeri, 20 to 30 percent of pseudomonas putida, 10 to 20 percent of pseudomonas-pertucinogena and 30 to 50 percent of bacillus subtilis. The compound fungus agent is prepared by combining and screening the strains, performing slant culture and primary culture, mixing according to the proportion, and performing secondary cycle culture. The preparation method is widely used in a biological enhancing water treatment process, has a long-term and stable operation period, and has specific degradation ability for toxic and harmful organic pollutants such as halogenated hydrocarbons, phenol, benzoic acid, aromatics, polycyclic aromatic hydrocarbon, heterocyclic compounds, phthalic acid ester and the like.

The fast tissue culture propagation process for azalea and camellia includes the following steps: cutting tender stem segment from the maternal plant in April or May; soaking in alcohol solution, sterilizing in HgCl2 solution and cutting the stem tip; inoculating the stem tip in primary culture medium to grow cespitose bud and cutting single bud seedling after the bud grows to 2-3cm length; culturing the single bud seedling in secondary culture medium inside a sealed container to proliferate test tube seedling repeatedly; culturing the test tube seedling in rooting culture medium inside sealed container to grow new root to 0.3-0.5cm long, and transplanting. The process can provide seedling of azalea and camellia continuously.

The invention discloses a benthic diatom culture method for growing seedlings and an abalone fry culture method, which relate to an abalone culture technology. The invention is characterized by comprising the following steps: (1) starting culturing algaes 45 days before pulling seedlings, keeping the culture water temperature at 14 DEG C and the illuminance between 20001 and 30001x, controlling the light between 10001 and 15001x at the last culture after screening and eliminating algae culture for one and half a month, and laying ovum after 7 days; (2) adding nutrient salt at primary culture,wherein the adding ratio of N:P:Si:Fe is 5:1:1:1(ppm); (3) laying ovums about 5 millions for each pool and carrying out micro inflation; (4) changing water regularly after the ovums are laid and changing water twice a measuring range which is 24 hours; (5) carrying out running water culture after the ovums swim for 5 to 6 days and are adhered; (6) controlling the illuminance within 10001x after larvas metamorphose; and (7) dumping the pool after treating copepods at about 30 days of the culture period, adjusting the illuminance between 15001 and 20001x and peeling off baby abalones after about45 days. The methods have the advantages of improving the yield and preventing and treating a plate stripping disease.

The invention relates to an exosome derived from human pluripotent stem cells, a preparation based on the exosome and use of the exosome and preparation, and belongs to the technical fields of cell biology, molecular biology and drug research and development. The human pluripotent stem cell exosome or the preparation based on the human pluripotent stem cell exosome has the uses of A, being an additive cultured in vitro for maintaining the stemness of adult stem cells and inhibiting the senility of the adult stem cells; B, being an additive cultured in vitro for maintaining the proliferative activity of various tissue primary culture cells; C, being applied in preparing medicine for treating bone and cartilage degenerative changes; D, being applied in preparing medicine for preventing the bone and cartilage degenerative changes; E, being applied in preparing medicine for treating orthopedic diseases such as osteonecrosis, bone nonunion, fractures, cartilage injuries, tendon injuries, frozen shoulders or tendon adhesion. Compared with the prior art, the exosome secreted by the pluripotent stem cells has powerful functions of promoting tissue cell regeneration, preventing senility andtreating bone diseases.

The invention relates to the technical field of adipose mesenchymal stem cells in the technical field of cell therapy, in particular to a method for enhancing an immune regulating function and a migration capacity of adipose mesenchymal stem cells. Primary culture and secondary culture are performed to the adipose mesenchymal stem cells obtained through separation, and then short-time culture is performed by using culture medium containing low-concentration TLR3 activator Poly(I:C). After Poly(I:C) co-stimulation culture is performed, the obtained adipose mesenchymal stem cells with a brand new phenotype have a stronger migration capacity, the levels of secreting cytokine and chemotactic factor are changed and a stronger immunosuppressive function is reflected on the whole.

The present invention provides a standardized tissue-specific and cell-specific kit and methods for promoting the enrichment and expansion of primary cells in culture while reducing the contamination of unwanted cell types. The present invention further provides the compositions for optimized tissue-specific and cell-type specific dissociation of tissues and inhibition of contaminating cell-types in primary cultures.

The invention provides a human brain glioma cell line and an establishing method and application thereof. The human brain glioma cell line is preserved in China Center for Typical Culture Collection with an accession number of CCTCC No. C201115. The human brain glioma cell line is established by subjecting brain glioma cells originated from a clinic specimen to primary culture and subculture, can be directly used for in vitro drug screening or generation of human brain glioma in mammals and is used for establishing a human brain glioma animal model and screening candidate drugs used for treating human brain glioma. The human brain glioma cell line has stable properties, can realize stable multiple passages and maintain stable properties even after in vitro passage to the 50th generation; pathogenesis of human brain glioma, drug susceptibility, transitivity and other related characteristics can be analyzed in vitro and in vivo, so two correlated in-vitro and in-vivo anti-brain glioma drug screening platforms can be established, and novel experimental materials closer to clinic oncobiological characteristics are provided for research on human brain glioma.

The invention discloses a complete medium and a human amnion-derived mesenchymal stem cell (hAMSCs) culture method. The complete medium is prepared by adding 3 to 10 percent of autologous umbilical cord blood serum into low-sugar Dulbecco minimum essential medium solution according to a volume ratio. The culture method comprises: (1) separation; (2) primary culture; and (3) subculture. The methodusing the complete medium in the hAMSCs culture has the advantages that: the risk of using fetal calf serum is avoided; although the need of adding L-glutamine, non-essential amino acid, 2-mercapitoethanol, pyruvic acid and the like is obviated, the high proliferation properties and phenotypic characteristics of the hAMSCs and expression of multilineage differentiation marker genes sand proteins of some stem cells can still be retained; and in subculture, the wall adherence fastness of the hAMSCs is much lower than that in fetal bovine serum (FBS) culture, the digestion time is reduced obviously, and the damage of trypsinization to cells and loss of cells are reduced.

The invention discloses a method for primary culture of tumor cells and relates to the field of cell biology. According to the method disclosed by the invention, a tumor sample is prepared into a single-cell suspension liquid by use of an enzyme method or a mechanical method, and then tumor cell bladders are prepared by use of a serum-free suspension culture method, wherein alkaline fibroblast growth factors, epidermal growth factors, insulin and bovine serum albumin need to be added during a culturing process, then the growth factors are removed, and the culture is implemented in a general culture medium to obtain adherent cells capable of realizing continuous passage. The method disclosed by the invention is short in culture period and has the effect of increasing the success rate of the primary culture of tumors.

The invention mainly relates to the field of plant tissue culture and particularly relates to a butterfly orchid anti-browning tissue culture method and an anti-browning culture medium. The tissue culture method comprises the steps of explant disinfection, primary culture, proliferation and differentiation culture, rooting culture, seedling hardening, transplanting and the like, wherein in the primary culture and the proliferation and differentiation culture, extracted solutions of calendulas, lemons, tomatoes, rosemary and the like are added into a corresponding culture medium so as to effectively prevent and treat a common browning problem in butterfly orchid tissue culture; and meanwhile, different types of cutting manners are used for helping to prevent and treat the browning problem at a differentiation phase, so as to form a compound anti-browning method. The method disclosed by the invention has the advantages of high stability, great success rate, greenness and environmental friendliness, and can annularly and repeatedly produce.

The invention discloses Lactobacillus-based mixed bacterium strains fermented minced fish. The finished product of the fish is prepared by a method comprising the steps of using minced fish as a base stock, inoculating the minced fish with mixed basic bacteria I having a colony density of 10 CFUs per gram of minced fish and mixed basic bacteria II having a colony density of 10 spores per gram of minced fish, placing the minced fished inoculated with primary cultures in a container for fermentation till the acidity reaches a standard degree, mixing the fermented mixture with a filling and adding the filling and subjecting the obtained mixture to vacuum degassing, preparation and packaging. The invention adopts a technical proposal that the minced fish is inoculated with primary cultures of the mixed basic bacteria I which are cultured by microorganic streptococcus lactis, lactobacillus bulgaricus and saccharomyces cerevisiae of the same type and have a colony density of 10 CFUs per gram of minced fish and the mixed basic bacteria II which are cultured by monascus and have a colony density of 10 spores per gram of minced fish for fermentation; the fermented mixture is mixed with a filling; the obtained mixture is added with the filling; and the final mixture is subjected to vacuum degassing and preparation. The method overcomes the drawback that natural fermentation cannot ensure the sanitation and safety, but leads to high salt content. The method can manufacture various finished products of fermented minced fish.

The invention discloses a composition for preventing browning of a plant tissue culture. The composition contains a direct antioxidant against phenolic substances and a plant antioxidase system reinforcing agent, and can play the systematic synergic action of two anti-browning mechanisms during the plant tissue culture process, on the one hand, the anti-oxidation action of the anti-oxidant can be realized against the phenolic substances infiltrating into a culture medium so as to reflect the direct reduction action, on the other hand, the plant antioxidase system reinforcing agent can reinforce the activity of antioxidase in strawberry tissues in plant tissues, and the production and the expansion of basic factors of the browning, namely quinone type substances can be reduced; and the combination of the two can effectively prevent the browning phenomenon in primary culture of strawberries and reduce the browning rate from the conventional level of above 30% to below 20%. The composition disclosed by the invention is widely suitable for various plants which are prone to browning in different tissue culture stages. The invention further discloses a using method of the composition.

The invention discloses a method for optimizing a potato isolated culture one-step seedling culture medium. The culture medium makes leaves of a potato test tube plantlet undergo isolated culture to form a seedling in one step. The one-step seedling culture medium takes an MS culture medium as a basal medium, and 6-benzyladenine, naphthyl acetic acid and 2,4-dichlorphenoxyacetic acid with different concentrations and combinations are supplied in 1L of the MS culture medium; adventitious buds and adventitious roots are directly differentiated through callus induction; and the potato regenerated seedling is obtained in one step. The callus obtained by induction in a primary culture medium is unnecessarily inoculated to a differential medium for regenerating the seedling. Compared with an isolated culture multi-step regenerated seedling method, the method of the invention has the advantages of simplified steps, short culture period, high repeatability, and high seedling survival rate.

The invention relates to an experimental method for analyzing the toxicity and the effect of a skin whitening agent through a primary culture human being skin melanocyte. The method comprises the following steps: 1, culturing and identifying the primary human being skin melanocyte; 2, carrying out the analysis of the toxicity of skin whitening substances through the primary human being skin melanocyte; and 3, carrying out the analysis of the effect of the skin whitening substances through the primary human being skin melanocyte, namely carrying out the analysis on two aspects of the melanin content and the tyrosinase activity. The method has the following advantages that: the human being melanocyte cultured in vitro has a high division and multiplication capacity, a high standardization degree, little difference between batches, and the same function of producing melanin granules with the melanocyte in vivo; the result obtained through the healthy human being melanocyte is more reliable than the result obtained through animal or human-derived melanoma cells; and the toxic action and the whitening effect of the substances to be tested are evaluated from the two aspects of characterization and quantification. The method can replace the living body animal and human being skin, and can be directly applied to toxicity and effect experiments of the whitening substances in products such as chemicals, cosmetics, medicaments, and the like.

The invention provides an establishing method of an immortal AM cell line, which establishes the immortal AM cell line by constructing a retrovirus carrier to infect cell activation telomerase. The establishing method comprises the following processes: constructing the pLXSNneo-hTERT retrovirus carrier, and carrying out AM cell primary culture and AM cell immune cell chemical identification to establish a stable and reliable immortal AM cell line. The cell line meets the features of epithelial cells, can ensure the stable expression of a target gene and provides a valuable in-vitro model for researching AM development and invasion features.

The invention provides a separation method for human amniotic mesenchymal stem cells (hAMSCs) in a process of cultivating the hAMSCs through enzyme digestion and epidermal growth factor (EGF). The cultivating process comprises the steps of separation of hAMSCs, primary culture of hAMSCs, hAMSCs subculturing and amplification, hAMSCs cryopreservation and recovery, and hAMSCs immunophenotyping detection. According to the method disclosed by the invention, step-by-step digestion is implemented for multiple times through trypsin and collagenase to collect amnion cells, and adherent culture and digestion time control are combined through adding of 4ng / ml of EGF (Epidermal Growth Factor), so as to obtain human amniotic mesenchymal stem cells (hAMSCs) which are relatively high in both purity and activity. The method disclosed by the invention, which combines enzyme digestion and EGF, has the advantages of being simple to operate, rapid, practical and the like, being an effective method for separating and purifying hAMSCs.

The invention belongs to the technical field of plant tissue culture and particularly relates to a method for building a bamboo reed tissue culture system. The method comprises the steps of (1) explant disinfection, wherein axillary buds of bamboo reeds are fully soaked in 75% of ethanol for 10 seconds and then moved into 0.2% of mercury bichloride solution for 10 minutes; (2) primary culture, wherein a culture medium is formed by adding cane sugar 30g / L and agar 8g / L in MS, then adding 6-BA 5mg / L and IBA 0.5mg / L and finally adjusting potential of hydrogen (pH) to 5.8; (3) secondary culture, wherein a culture medium and culture conditions of the secondary culture are the same as the culture medium and culture conditions for the primary culture; (4) strong bud and root culture, wherein the buds are cultured in the secondary culture medium for one month to finish strong bud culture and then planted in a rooting culture medium, and the rooting culture medium is formed by adding IBA 0.5 mg / L into the MS; and (5) domestication and transplantation, wherein the domestication is performed by using a floating bed domestication system for one week and then transplantation is performed. By means of the method, the efficient breeding of the bamboo reeds can be achieved.

A method of identifying a micro-organism and determining the susceptibility of the micro-organism to an antimicrobial agent by inserting a sample of the micro-organism in a primary culture container and allowing the micro-organism to multiply. The primary culture is then divided into a plurality of secondary cultures and the metabolic volatile or semi-volatile compounds (VCs) in the headspace above the cultures are analysed by SIFT-MS to ascertain whether micro-organisms exist in the culture and determine the type of micro-organism. The secondary cultures are then divided into a number of tertiary cultures and an antimicrobial agent is introduced whereupon the VCs in the headspace above the tertiary cultures are analysed by SIFT-MS to determine the susceptibility of the micro-organism to the antimicrobial agent at various concentrations of the antimicrobial agent.

The invention discloses a cell line of pterygiophore tissue of cryprinus carpiod and a construction method. The construction method comprises the following steps of: A, preparing pterygiophore tissue blocks of cryprinus carpiod, namely putting cryprinus carpiod into potassium permanganate solution, disinfecting, transferring pterygiophore tissue to a culture dish, cleaning with an antibiotic, and transplanting the pterygiophore tissue blocks in cell culture flasks; B, preparing proliferation culture solution special for cells of pterygiophore tissue of cryprinus carpiod, namely adding fetal bovine serum, adding alkaline fibroblast growth factors and epidermal growth factors into the culture solution; C, performing primary culture of the cells of pterygiophore tissue of cryprinus carpiod, namely adding the special proliferation culture solution into pterygiophore tissue which is subjected to dry sticking in each culture flask, culturing in the culture box, and adding the proliferation culture solution special for the cells of pterygiophore tissue of cryprinus carpiod; and D, performing the subculture of the cells of pterygiophore tissue of cryprinus carpiod, namely after the cells of pterygiophore tissue of cryprinus carpiod are grown to form a single layer, preparing cell suspension, and performing the subculture. The method is easy and convenient to operate and has a scientific and reasonable process, and the established cell line of pterygiophore tissue of cryprinus carpiod is subcultured for over 60 generations at present.

A nanostructured molecular delivery vehicle comprising magnetic materials and configured to receive passenger biomolecules. The application of a an appropriate magnetic field having a gradient orients and drives the vehicle into a biological target, which may comprise cells, cell masses, tissue slices, tissues, etc. Under the control of the magnetic field, these vehicles can penetrate cell membranes. Then, the biomolecules carried by the vehicle can be released into the cells to perform their functions. Using this “nanospearing” technique, unprecendented high transfection efficiency has been achieved in several difficult-to-transfect cells. These include, but are not limited to, Bal 17 cells, ex vivo B cells, primary cultured cortical neurons, etc. This method advances the state of the art, providing an improved technique for the introduction of exogenous molecules to cells, with the clinical applications including, but not being limited to, drug delivery, gene therapy, vaccination, etc.

A tissue culture method for new pteris fern includes such steps as obtain aseptic material, primary culture with the culture medium containing MS, 6-BA, NAA and AC, reproduction culture with the culture medium containing MS, 6-BA and AC, rooting culture with the culture medium containing 1 / 2 MS, NAA and IB, and intelligent culture of seedlings.

The invention provides a method of producing pearls on the basis of pearl shell mantel cell culture. The method includes: cutting mantel of a pearl shell into small slices, scattering to obtain single cells by enzymatic depolymerization, performing primary culture prior to subculturing, centrifuging, preparing solution by obtained cells, quantitatively injecting connective tissue in pearl shell mantel with the solution, placing the pearl shells in a culture area, and culturing for one cycle to obtain pearls.

The invention provides a method for tissue culture of paeonia lactiflora, which comprises the following steps of: sterilizing a paeonia lactiflora stem tip; and performing primary culture, secondary culture and rooting culture on the sterilized paeonia lactiflora stem tip, wherein the sterilization of the paeonia lactiflora stem tip is implemented by two steps to ensure that the stem tip is sterilized thoroughly, an explant is not damaged, and the survival rate of the stem tip is high. The method for tissue culture of the paeonia lactiflora is simple to operate, has a low cost and a high rooting rate, can realize transplantation out of a bottle, and is suitable for rapid propagation of most ornamental paeonia lactiflora varieties; and simultaneously, according to the culture method provided by the invention, the excellent characteristics of an original species can be kept, and a base is established for high-quality seed breeding and industrial production.