41results about How to "Promotes adherent growth" patented technology

The invention discloses a mesenchymal stem cell serum-free culture medium. The mesenchymal stem cell serum-free culture medium comprises a basic culture medium and added ingredients added in the basic culture medium, and the added ingredients comprise L-glutamine, nonessential amino acid, L-ascorbic acid, sodium selenite, fibronectin, ethanolamine, hydrocortisone, a trypsin inhibitor, human transferrin, human insulin, bFGF, TGF-beta1 and PDGF-BB. By means of the culture medium, the problems that in the prior art, the serum-free culture medium is poor in cell adherence, components are relatively complex, and primary cell culture is not supported are solved.

The invention discloses a chitosan polyacrylamide hydrogel base material and a preparation method thereof. The chitosan polyacrylamide hydrogel base material is composed of an acrylamide solution, a methylene diacrylamide solution, H2O, an ammonium persulfate solution, tetramethyl ethylenediamine and a chitosan solution. The preparation method of the chitosan polyacrylamide hydrogel base material comprises a step of mixing and stirring, a step of transferring to a glass clamping plate for reacting, and a step of immersion. The chitosan polyacrylamide hydrogel base material has a simple technology and allows hydrogels having different hardnesses to be obtained, and the obtained hydrogels have the advantages of non-toxicity, easy adherence growth of cells, reduction of the reaching complexity of the cytomechanics, benefiting for the researches on the mechanical properties of the cells, and reduction of the researching cost.

The invention discloses collagen-based composite hemostasis powder and a preparation method thereof. The method is characterized by comprising the following steps: modifying type I collagen by using dencichine, preparing the modified type I collagen into a solution with the concentration of 1-2 percent by using polylysine-modified chitosan; preparing medical gelatin into a 1-3% solution; preparing the modified chitosan into a 1-3% solution; blending the three solutions according to a mass ratio of 0.5-1:2-8:8-2; stirring at the temperature of 4-10 DEG C for 8-10 hours, and performing freeze drying to prepare composite sponge; sequentially impregnating the composite sponge in a modifier and a blood coagulation factor in an ultrasonic cleaner, fully washing, performing freeze drying, and grinding by a grinder to prepare the collagen-based composite hemostasis powder. The powder has the effects of rapidly and effectively stopping bleeding, resisting bacteria, diminishing inflammation, relieving pain, promoting wound healing and repairing, has high adhesion property, hydrophilicity, biological degradability and biocompatibility and is low in price and suitable for wound hemostasis or repair.

The invention relates to a preparation method of composite cavity microfibers based on a micro-fluidic technology. The preparation method is characterized in that in the preparation process of cavitymicrofibers, a modified material capable of promoting the cell wall adhesion growth is introduced in a microfiber inner cavity; while the cavity is formed, the modified material is adhered onto the cavity to form a modified coating layer, so that a promoting effect on late cell adhesion and culture is provided. By utilizing the micro-fluidic technology, a micron-sized channel capable of generatinga coaxial laminar flow pattern is formed, the flow pattern control on a sample fluid is realized, and the sample fluid is finally solidified into a micron-sized hollow fiber material with a specificinner coating structure. The microfiber material can simulate a microstructure in a human body tissue, and a new method and a new concept are provided for tissue engineering and organ regeneration. The preparation method provided by the invention is simple and reliable to operate, high in efficiency and excellent in technical effect; convenient conditions are provided for the modification of the microfibers; an internal finish coating is uniform, stable, simple and controllable to facilitate the cell wall adhesion growth.

The invention discloses a manufacturing method for a hard micro-fluid chip. The manufacturing method comprises the following steps: preparing an upper chip and chemically bonding and packaging the upper chip under a semi-cured state with a substrate, thereby acquiring the hard micro-fluid chip. The upper chip can be prepared from epoxy resin or amino resin; when the upper chip is prepared, the heating curing temperature is 45-85 DEG C and the time is 15 minutes to 8 hours; a thermal polymerized epoxy resin material is used after the two materials of prepolymer and curing agent are mixed, the viscosity thereof after mixing is low (approximate to mineral oil viscosity), the rolling printing for a tiny structure at micron scale and even nanometer scale is convenient and the rolling precision is high; the mixture can be solidified within 40min under the temperature of 80 DEG C, the preparation time of the micro-fluid chip is shortened, the period is short, the consumption of a reagent is less and the batch production is convenient; the hard micro-fluid chip can be flexibly packaged and combined with various substrates; the packaging is independent of external high temperature and high pressure environments; and the packaging strength is high and the speed is high.

The invention belongs to the field of bioengineering, and more specifically relates to a method for preparing serotype 8 recombinant adeno-associated virus, The method comprises the following steps: 1) using a serum-free medium for culturing 293T cell in a sheet carrier; 2) performing transient transfection on the cultured 293T cell by adeno-associated viral vector system plasmid to obtain the transfected cell; and the adeno-associated viral vector system plasmid comprises AAV expression vector plasmid, auxiliary plasmid pHelper and AAV capsid plasmid pRC8; and 3) continuously culturing the transfected cell, collecting and replacing the serum-free medium on time to obtain a medium containing the recombinant adeno-associated virus. The preparation method can guarantee production stability,preparation and purifying steps are saved, downstream virus purifying difficulty is obliviously reduced, and the preparation method is especially suitable for industrial scale-production of the serotype 8 recombinant adeno-associated virus.

The invention discloses a bioreactor for in vitro culture of neurotrophic factors, which comprises an upper cover and a lower cover which are provided with concave cavities. The concave cavities of the upper cover and the lower cover are closed in opposite directions and fixed to form a box body, a culture plate through which cerebrospinal fluid can pass is arranged in the cavity of the box body,a polyether sulfone semi-permeable membrane is covered on the culture plate, the culture plate is embedded to the middle lower part of the concave cavity of the lower cover and positioned between theupper cover and the lower cover, both the upper part and the lower part of the culture plate are spaces in which the cerebrospinal fluid flows, the upper cover is provided with an input nozzle, the lower cover is provided with an output nozzle, and the upper end face of the upper cover is provided with a plurality of holes through which the air enters the box body. The bioreactor enables the cerebrospinal fluid of a nervous system dysfunction patient to be circulated in the bioreactor and fully mixed with the neurotrophic factors secreted by star-shaped gliocytes to form mixed cerebrospinal fluid containing the neurotrophic factors to be redelivered to the lower cavity of the arachnoid membrane or the ventricular system of the patient, thereby realizing promotion of recovery of functionaldeficiency or functional disorder of the central nervous system caused by diseases or injury, and remarkably improving the life quality of the patient.

The invention discloses a primary culture method of the embryonic cell tissue pieces of a Chinese softshell turtle. The primary culture method comprises the following steps of: (1) collecting the fertile eggs of the Chinese softshell turtle, and disinfecting; (2) breaking the eggshells of the fertile eggs, separating embryos from yolk, cleaning the separated embryonic tissues with a 1*PBS (Phosphate Buffer Solution), and then soaking for sterilization; (3) cleaning the embryonic tissues for 4-5 times in an M199 culture medium, then transferring into a complete culture medium, cutting the embryonic tissues into pieces to obtain tissue pieces, and then cleaning the tissue pieces once by using the complete culture medium; (4) uniformly dispersing the tissue pieces to the inner wall of a cell bottle by using a sterile disposable inoculating needle so that the tissue pieces grow for 3-4 hours in a way of being attached to the wall, then adding the complete culture medium to submerge the tissue pieces, and culturing at the constant temperature of 30-31 DEG C for more than 3 days. The primary culture method disclosed by the invention has the advantages of easy operation, convenient culture, no need of specific cell separation liquid and high culture success rate.

The invention belongs to the field of cell culture and particularly relates to a low-serum protein-free culture medium applicable to Marc-145 cell growth and a preparation method thereof. The culture medium comprises amino acid, vitamin, inorganic salt, micro elements and other ingredients. The low-serum protein-free culture medium applicable to Marc-145 cell growth and the preparation method of the low-serum protein-free culture medium have the advantages of being free of protein and animal source ingredients and definite in chemical ingredient, the interference to cell culture of serum is industrially reduced, the production cost is lowered, and great convenience is brought to industrial production of cells.

The invention provides a method for preparing dental pulp stem cells. The method comprises the following steps of S1, collecting teeth; S2, separating the dental pulp stem cells, fixing the teeth, splitting the dental crown, fetching out dental pulp, putting the fetched dental pulp into PBS (phosphate buffer saline) for flushing, then preparing the fetched dental pulp into blocks with size of 1 to3 mm<3>, adding a mixed solution of 0.025% to 0.2% of I-type collagenase and 0.025% to 0.2% of neutral protease, digesting for 10 to 30 min at the temperature of 37 DEG C, adding the PBS, centrifuging at the speed of 1000 to 1500 rpm to remove supernatant, settling, adding a culture medium for moistening, attaching the tissue blocks into a cell culture dish, and putting into a CO2 (carbon dioxide) culture box to culture. By adopting the technical scheme, compared with the prior art, the method has the advantages that the enzyme used by the digestion wall-attaching method is a mixture of the I-type collagenase and the neutral protease, and the functions of the I-type collagenase and the neutral protease are mild, so that the restriction of surrounding tissues to the stem cells is reduced,and the damage to the cells is avoided; the purity of the separated stem cells is guaranteed, and the cell harvesting time is shortened.

The invention discloses a decidua parietalis mesenchymal stem cell culture medium for treating tumors and a co-culture method. The decidua parietalis mesenchymal stem cell culture medium for treating tumors contains decidua parietalis mesenchymal stem cells and a selective culture medium, the decidua parietalis mesenchymal stem cells have a homing effect, a proliferation inhibiting effect and a cell cycle adjusting effect on various tumor cells.

The invention relates to a permeable protection agent, a serum-free frozen stock solution, and preparation and application methods of the serum-free frozen stock solution. The permeable protection agent is fulvic acid. The technical scheme of the invention has the technical effects that the cost is low; the toxicity is low; and the survival rate of cells is high after resuscitation.

The invention relates to a method for producing a porcine pseudorabies gE gene deletion virus vaccines by using a BHK cell line. The method comprises the following steps: resuscitating BHK21 cells and performing micro-carrier culture by adopting a low-serum culture medium; then replacing the low-serum culture medium with a cell maintenance solution, performing continuous perfusion culture, inoculating when the cell density is maintained at (1.5-3)*10<7> pcs/ml and continuously harvesting virus liquid for six days after 48 hours; concentrating, inactivating and sterilizing the harvested virus liquid to prepare the finished product vaccine. The method provided by the invention has the advantages that the vaccine is high in potency and stronger in mmunogenicity and can achieve a good immune effect without adding an immunoenhancer and promote the secretion of neutralizing antibodies in vivo after immunization; the immune protective rate achieves 100 percent, so that the vaccine efficacy evaluation criteria is fully reached.

The invention discloses a method for preparing feed layer cells by using R6 transfected MEF. The method comprises the following steps: firstly using suppression transcription factor R6 transfected MEFbased on transcriptional activation effect material sample Tale combined inside an Xist first intron area to obtain immortal type MEF cell lines R6-MEF, after screening the various R6-MEF cell lines,selecting the R6-MEF cell lines with relatively high multipotential gene expression, and after mitomycin treatment, obtaining the novel feed layer cells R6-feeder, so that the obtained feed layer cells can be preferably used for maintaining the pluripotency iPSC of embryonic stem cells ESC and induced multipotential stem cells. According to the method, a new selection is provided for cultivationand research of stem cells, and a new technological and theoretical basis is provided for researching how to obtain and improve iPSCs of people, pigs, cows and other mammals.

The invention discloses a preparation method of mesenchymal stem cells derived from early mesoderm. The method comprises the steps of cleaning, enzymolysis, filtering, primary culture, subculture, induced differentiation and induced differentiation. The preparation method has the beneficial effects that the separation efficiency of the stem cells is improved, so that temperature change during heating and refrigeration of the stem cells becomes more smooth, cell inactivation caused by the temperature change in a cell extraction process is avoided, the problem of the extraction rate of the cellsis reduced, the separation speed of the mesenchymal stem cells can be improved, purity of the mesenchymal stem cells in a separation process is ensured by polylysine, and growth of the cells adheringto walls after separation is promoted.

The invention provides a super-hydrophilic coating layer for cell culture and a preparation method. According to the super-hydrophilic coating layer for cell culture and the preparation method, the polyvinyl alcohol-chitosan interpenetrating network super-hydrophilic coating layer is coated on a cell culture device in a spraying, dip-coating or roll-coating manner and the like, the super-hydrophilic coating layer has a durable and stable super-hydrophilic effect, and the water contact angle is smaller than 10 degrees; the prepared coating layer of the cell culture device is strong in adhesiveforce, high in transparency, resistant to ultraviolet and water soaking, good in biocompatibility, low in cytotoxicity and high in cell adherence rate.

The invention relates to the technical field of cell cryopreservation, and concretely relates to serum-free cell cryopreservation liquid, and a preparation method and a preparation device thereof. The cryopreservation liquid comprisesa DMEM/F-12 basic culture medium, Nutridoma, pyruvic acid, L-glutamine and 0.1-0.41 g/L of an intracellular and extracellular cryoprotectant. The cell cryopreservation liquid without serum or DMSO is determined in formula components and has very high biological safety and clinical application prospects, the recovery survival rate of cells cryopreserved with the cell cryopreservation liquid reaches 87% or above, good cell viability and physiological characteristics can be maintained, and the cryopreservation liquidis very suitable for the related research field of primary cells and stem cells.