845 results about "Fetal bovine serum" patented technology

The invention discloses a method for constructing a human peripheral blood immune cell bank. The method comprises the following steps of: collecting human peripheral blood, separating autologous plasma, separating a peripheral blood mononuclear cell, separating a mononuclear cell by the peripheral blood mononuclear cell and freezing, separating a T lymphocyte by the peripheral blood mononuclear cell and freezing, separating a B lymphocyte by the peripheral blood mononuclear cell and freezing, separating an NK cell by the peripheral blood mononuclear cell and freezing, and encoding and puttingin storage. According to the invention, immune cells of health or young people are separated and are respectively independently frozen, and relative numbers are stored and put into storage, so that the stored human immune cells have the characteristics of high activity, high purity and convenience for use, and fetal calf serum can be replaced by the human autologous plasma, so that introduction of a foreign protein can be avoided. Meanwhile, the method, provided by the invention, has the advantages of low cost, low requirements on laboratory conditions, and wide application.

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.

A cryopreservation solution of tissue engineering products uses DMEM culture solution as a basic solvent which is added with vitamin B, vitamin C, chondroitin sulfate, beta-integrin, cromolyn sodium, cytochalasin B, L-glutamine, bovine serum albumin, fetal bovine serum, trehalose, sodium carbonate, polysucrose-70, and dimethyl sulfoxide added during freezing storage. The cryopreservation solution provided by the invention has little toxicity to cells and long storage time, and can be stored for six months under 80 DEG C below zero, and 12 to 18 months in liquid nitrogen; and the cell viability of the resuscitated cells can be over 60%. The stored tissues can be simply and conveniently used after being resuscitated for only three to five min in water bath under the temperature of 37 DEG C and being washed by sterilized saline water. The invention can be widely used, and is suitable for tissue engineering skin, tissue engineering cornea, tissue engineering blood vessel, tissue engineering nerve and the like, and also is applicable to the cryopreservation of normal tissues.

The invention provides a culture fluid exclusively used in cow in-vitro fertilization embryos. The culture fluid contains 109.0mM-110mM of NaCl, 2.9mM-3.1mM of KCl, 26.0mM-26.5mM of NaHCO3, 0.5mM-1.0mM of MgCl2.6H2O, 1.0mM-1.3mM of KH2PO3, 0.4mM of sodium pyruvate, 1.5 mM of glucose, 5mM of galacturonic calcium, 10v/v% fetal bovine serum, 1mM of L- glutamine, 2v/v% essential amino-acid, 1v/v% nonessential amino acid and 1mM-10mM of glutathione. The cow in-vitro fertilization embryos are placed into the culture fluid to carry out in-vitro cultivation, and results are shown to be obviously superior to those of a control group which is not added with GSH (glutathione), and therefore, developmental rate of blastula and embryo quality are improved, cost for producing embryos in vitro is lowered, experimental basis is provided for applying a cow IVF (in vitro fertilization) technology to practice, and a genetic breeding process can be greatly accelerated.

The invention relates to a cell cryopreservation liquid and a cell cryopreservation method. The cell cryopreservation liquid comprises 90-95% of a whole cell culture liquid and 5-10% of dimethyl sulfoxide according to the volume percentage, wherein the whole cell culture liquid contains a cell culture medium and fetal calf serum with the volume ratio of 9:1. The whole cell culture liquid of the cell cryopreservation liquid provides essential nutrient substances for cellular life metabolism, and the dimethyl sulfoxide is used as an antifreezing agent; through reasonable proportion of the amounts of the cell culture medium and the fetal calf serum in the whole cell culture liquid and the amounts of the whole cell culture liquid and the antifreezing agent, the whole cell culture liquid with lower content of the fetal calf serum can provide the essential and enough nutrient substances for the cellular life metabolism; moreover, the appropriate amount of the antifreezing agent can prevent or reduce damage effects of frozen ice crystals on cells so as to improve the survival rate of the cells; and the cell cryopreservation liquid contains no serum easily causing pollution, thereby having lower pollution risk.

The present invention relates generally to nutritive (e.g., cell culture) medium, medium supplement, media subgroup and buffer formulations. Specifically, the present invention provides powdered nutritive medium, medium supplement and medium subgroup formulations, particularly cell culture medium supplements (including powdered sera such as powdered fetal bovine serum (FBS)), medium subgroup formulations and cell culture media comprising necessary nutritive factors that facilitate the in vitro cultivation of cells. The invention further provides such powdered formulations that produce particular or desired final ionic and/or pH conditions upon reconstitution with a solvent. The invention is also directed to methods of production of these formulations, and also provides kits and methods for cultivation of prokaryotic and eukaryotic cells using these formulations. The invention also relates to methods of producing sterile formulations, and to methods for producing dry cell powders. The invention also provides cell, media, media supplement, media subgroup and buffer powders produced by the methods of the invention.

The invention discloses a method for preparing human umbilical cord mesenchymal stem cell exosomes. The method uses a newborn umbilical cord as a source of umbilical cord mesenchymal stem cells, usesa medium prepared by a fetal calf serum or a serum substitute in which exosome carried by itself is removedfor culturing, collects a supernatant, and further separates and extracts the exosomesby differential centrifugation, the purity is high, and the source of the umbilical cord mesenchymal stem cell exosomes is ensured; the exosomes is prepared by the differential centrifugation, the operationis simple, and the method is suitable for large-scale production; during the whole process, sterility is guaranteed, the contamination ofmycoplasma is prevented, and the product safety is higher.

The invention discloses a stem cell medium and an application thereof. The medium comprises a basic medium, fetal bovine serum, cytokine or protein polypeptide, vitamin and lipid. The medium provided by the invention can quickly amplify stem cells and does not influence the potential of the stem cells, and the amplification velocity of the stem cells is improved by 3-5 times than a conventional medium; and moreover, the medium can be used for culturing the stem cells of various tissues and has excellent applicability, the cultured stem cells have strong differentiation capability and can be differentiated into cells with various functions; therefore, the stem cell medium has high scientific and research and medical application values.

The present invention provides an adipose tissue-derived mesenchymal stem cell amplification culture method, which comprises: preparing a suspension containing adipose tissue-derived mesenchymal stem cells with an adipose-derived stem cell culture medium, and inoculating the suspension into a stem cell culture device being subjected to a fibronectin coating treatment to carry out amplification culture. According to the present invention, the solid-phase support for growth and adhesion of the fibronectin-coated adipose-derived mesenchymal stem cells is used, and the synergetic screening culture of the low-concentration fetal bovine serum and the high-concentration gentamicin is used, such that the adipose tissue-derived mesenchymal stem cells can be amplified by 400-500 times within two weeks, the purity is high, and the adipose tissue-derived mesenchymal stem cells can be subjected to directional induction differentiation to obtain the adipose tissue.

The invention relates to a culture medium for a normal epithelial cell of a human or mammal, primary separation culture, subculture, 3D gas-liquid culture and 3D matrigel culture methods, the normal epithelial cell generated by using the culture medium and the culture methods and application of the normal epithelial cell to a toxicological evaluation system. The culture medium is prepared by mixing DMEM and Ham's F-12NUTRIENT MIX according to the volume ratio of 3:1 and also adding 4-6% of fetal calf serum, 1-3nM triiodothyronine, 0.4-0.65% of insulin-transferrin-selenium reagent, 4-6mu g/ml transferrin, 9-11ng/mL epidermal growth factors, 0.3-0.5mu g/mL hydrocortisone, 0.5-1.5nM cholera toxin, 0.4-0.6mu g/mL amphoterrible B, 35-45mu g/mL gentamicin, 45-55nM calpeptin, 35-45ng/ml recombinant human IL-1RA and 3mu g/ml recombinant human R-Spondin-1. The culture medium disclosed by the invention can be used for carrying out separation culture or subculture on the normal epithelial cell of the human or the mammal and any other various tissue source, rapidly proliferating the normal epithelial cell in vitro and establishing a cell line; and the normal epithelial cell is a normal diploid cell and is applied to the toxicological evaluation system of the human or the mammal.

Providing is cellular aggregates derived from corneal endothelial cells that, when transplanted, readily adhere to the parenchyma of cornea and function in a manner equivalent to corneal endothelial cells, and a method of transplantation of the cellular aggregates. Cellular aggregates derived from corneal endothelial cells. The cellular aggregates derived from corneal endothelial cells is prepared by culturing human corneal endothelial cells in a medium containing fetal bovine serum, growth factor and glucose; and then float culturing the cells obtained in a medium containing growth factor. A method of transplantation into the anterior chamber the cellular aggregate or the cellular aggregate prepared by the above method, comprising inserting a tube into the parenchyma of cornea, introducing the cellular aggregate into the anterior chamber through the inserted tube, and causing the cellular aggregate that has been introduced to adhere to Descemet's membrane by assuming in a downward-facing position.

Genes encoding the transcription factors controlling the core circadian oscillator (BMAL, Clock, NPAS, Per) and their regulatory targets (Rev-erbα, Rev-erb) have been found in adipose tissue. The circadian pattern of these genes was entrained using restricted feeding. The circadian gene expression profiles were examined in mice and in undifferentiated and adipocyte-differentiated human adipose stem cells following exposure to nuclear hormone receptor ligands (dexamethasone or thiazolidinedione) or 30% fetal bovine serum. All three agents induced the initiation of a cyclic expression profile in representative circadian genes in the human adipose stem cells. The circadian genes studied displayed an oscillatory expression profile, characterized by both a zenith and nadir within a 24-28 hr phase. The circadian gene pattern has been lengthened with use of an inhibitor of glycogen synthase kinase 3 beta. Modulation of the circadian pattern to lengthen or shorten can be used to affect weight gain or loss, respectively.

A process for generating megacaryocyte by in vitro induction to umbilical blood cell CD34+ includes in vitro amplifying the CD34+ in the culture medium containing fetal calf serum, human SCF, human TPO and/or ligand flt-3 and inducing the generation of megacaryocytes in the non-serum culture medium containing human TPO, human interleukin 3(IL-3) and/or human GM-CSF. Its advantage is high inducing efficiency.

The invention provides a clinic freeze-preservation protective solution composition for umbilical cord mesenchymal stem cells and application thereof. The protective solution composition comprises the following components: compound dextranum 40 injection, human serum albumin and DMSO. Compared with the prior art, the composition has the beneficial effects that the clinic freeze-preservation protective solution for umbilical cord mesenchymal stem cells can main the activity and biology activity of the mesenchymal stem cell for a long time, and does not contain unsafe components such as cell culture mediums, fetal calf serum and human plasma. All ingredients comprise clinical drug compound dextranum 40 injection, 20% human serum albumin or clinically used (dimethyl sulfoxide (UPS clinical level)), and the stability of the clinically standardized mesenchymal stem cell freeze-preservation system can be guaranteed while the umbilical cord mesenchymal stem cells for cryopreservation resuscitation meet the clinic using requirement. The application is safe, effective and simple in preparation method, and has good clinical application prospect.

The invention discloses a method for separately culturing a human adipose mesenchymal stem cell and a dedicated culture medium thereof. The culture medium used for separately culturing the human adipose mesenchymal stem cell comprises an animal cell basic culture medium, fetal calf serum, an epidermal growth factor and a platelet-derived growth factor. The final concentration of the fetal calf serum is 1-200 mL/L, the final concentration of the epidermal growth factor is 1-100 ng/ml, and the final concentration of the platelet-derived growth factor is 1-100 ng/ml. The adipose mesenchymal stem cell of the invention has CD31-, CD34-, CD45- and HLA-DR-, as well as the phenotype of CD29+, CD44+, CD105+ and Flk-1+. The specificity cell surface marker and the relevant antihelion molecule of a skeletal muscle cell and a vascular endothelia cell can be expressed after inducement is performed in vitro. Muscle fiber, vascular endothelin and functional muscle satellite cells can be differentiated in a muscle injury model mouse body caused by medicine and the expression of dystrophin protein on the ducheme muscular dystrophy (DMD) model mouse (mdx) myolemma can be partially recovered, so as to release the pathological symptom of the model mouse.

The invention discloses cell frozen stock solution for freezing and storing nerve cells and a freezing storage method, wherein, the cell frozen stock solution and the freezing storage method are used for freezing and storing the nerve cells, especially primary animal nerve cells that are isolated freshly, such as hippocampus nerve cells. The frozen stock solution is formed by adding basic medium,fetal bovine serum, dimethyl sulfoxide,6-fructose phosphate and flavonoid compounds to the basic medium; 6-fructose phosphate (w/w) achieves the function of improving cell viability (effective cell rate) when the frozen cells are recovered; and in addition, the flavonoid compounds achieve definite oxidation resistance and can prevent free radical from damaging the cells. The cell frozen stock solution and the freezing storage method can improve the survival rate and the cell function of the nerve cells through freezing storage, the survival rate of unfreezing cells can be increased to reach more than 95 percent, the frozen stock solution for the nerve cells can preserve the nerve cells for a long period, the biological activity of the nerve cells can be ensured, and very high utility values can be achieved.

The invention provides a kit and method for inducing differentiation from bone mesenchymal stem cells to osteoblasts. The kit comprises an osteoblast inducing culture solution and an osteoblast identification solution, wherein the inducing culture solution comprises a cell culture solution A, a cell culture solution B, a cell culture solution C, a cell culture solution D and a cell culture solution E; the osteoblast identification solution comprises a cell immobilizing solution, coloring agents and a detergent; the cell culture solution A is an alpha-minimum essential medium (MEM) liquid culture medium; the cell culture solution B is fetal bovine serum; the cell culture solution C is a dexamethasone solution; the cell culture solution D is a beta-sodium glycerophosphate solution; the cell culture solution E is a vitamin C solution; the cell immobilizing solution is 4% paraformaldehyde; the coloring agents include sodium thiosulfate, silver nitrate and neutral red; and the detergent is double distilled water.

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.

The invention relates to the technical field of biological control of breast cancer disease and discloses a polypeptide for promoting apoptosis of breast cancer cells by targeted uptake of siRNA. Thepolypeptide comprises a polypeptide 1; the sequence of the polypeptide 1 is H-Ile-Phe-D-Trp-Leu-Leu-Trp-Gln-Gly-Arg-Gly-Gly-Gly-Arg-Arg-Arg-Arg-Arg-Arg-Arg-OH. A detection method for the polypeptide for promoting the apoptosis of the breast cancer cells by targeted uptake of the siRNA comprises the following steps: firstly, culturing the breast cancer cells: selecting breast cancer cells MDA-MB-231, culturing the breast cancer cells MDA-MB-231 by adopting a DMEM culture medium, and sequentially adding 10 percent of fetal calf serum, 100 unit/ml of penicillin and 100g/mL of streptomycin in theculture medium. According to the polypeptide for promoting the apoptosis of the breast cancer cells by targeted uptake of the siRNA, the polypeptide 1 wraps the siRNA to form nanoparticles for targeting delivery of the siRNA to the breast cancer cells; the nanoparticles can realize the targeting delivery of the siRNA through a surface receptor of the breast cancer cells and have little damage to normal cells, and the practicality of the polypeptide is improved; meanwhile, the targeting delivery of TRPC1 siRNA by using the polypeptide 1 causes the apoptosis of the breast cancer cells, so that the effect of treating breast cancer is realized.

A medium for culturing stem cell. The stem cell medium of the invention comprises a fetal bovine serum, one or plurality of amino acid, one or plurality of vitamin, one or plurality of growth factor, one or plurality of inorganic salt, one or plurality of antioxidant, wherein the stem cell medium has a calcium concentration of less than about 1.8 mM, and the fetal bovine serum is present in an amount of less than about 10% by volume of the medium. The stem cell medium of the invention can maintain the proliferative and self-renewal capacity of the stem cells and keep stem cells at a steady stage.