111 results about "Transferrin" patented technology

The present disclosure is directed to bifunctional small molecules which contain a circulating protein binding moiety (CPBM) linked through a linker group to a cellular receptor binding moiety (CRBM) which is a membrane receptor of degrading cell such as a hepatocyte or other degrading cell. In certain embodiments, the (CRBM) is a moiety which binds to asialoglycoprotein receptor (an asialoglycoprotein receptor binding moiety, or ASGPRBM) of a hepatocyte. In additional embodiments, the (CRBM) is a moiety which binds to a receptor of other cells which can degrade proteins, such as a LRP1, LDLR, FcγRI, FcRN, Transferrin or Macrophage Scavenger receptor.

The invention relates to an isolated culture method of functional Chinese softshell turtle ovarian granular cells. The isolated culture method comprises the following steps: S1, collecting ovarian tissues: collecting Chinese softshell turtle ovarian tissues; s2, separating and collecting a granular cell layer; s3, digesting and collecting granular cells; s4, primary culture of granulosa cells: transferring the collected cells into a cell culture bottle containing 4 ml of a complete culture medium for primary culture, wherein the complete culture medium comprises the following main components: a DMEM culture medium, 15% of fetal calf serum, 1% of penicillin streptomycin and 1% of an insulin-transferrin-selenium additive; and S5, subculturing the granular cells. The isolated culture method and the culture medium system provided by the invention are simpler and more time-saving and labor-saving; according to the method, the aromatase CYP19A1 can be expressed normally, the characteristics of the granular cells are kept, the aromatase CYP19A1 can be expressed normally, it is indicated that the in-vitro cultured cells can keep the functionality of the cells, and the in-vitro proliferation capacity of the granular cells is improved to a great extent.

This invention discloses a NK cell culture kit, its culture method, and its application. The kit includes an amplification medium, a high-efficiency induction medium, an NK-A coating solution, an NK-B mixture, and an NK-C mixture. The amplification medium contains basal medium, NAD+, human serum albumin, transferrin, β-glucan, glutathione, β-mercaptoethanol, and linoleic acid. The high-efficiency induction medium, in addition to the amplification medium components, contains soybean peptides, nicotinamide, inulin, and N-acetyl-L-cysteine. The NK-A coating solution contains heparin sodium, monoclonal antibodies, and antibodies. The NK-B mixture contains Inbakicept, IL-2, and IL-15. The NK-C mixture contains linolenic acid, IL-2, and IL-18. The Inbakicept factor in the kit can activate NK cells and enhance their cytotoxicity; linolenic acid can inhibit T cell proliferation, increase NK cell purity, and enhance cell efficacy.

The invention belongs to the technical field of molecular biology, and relates to an application of transferrin (TF) as an internal reference protein in preparation of a reagent for detecting exosome proteins of neurodegenerative diseases. The TF can maintain a stable expression level in nerve cells, brain tissues, blood plasma and serum-derived exosomes of a subject suffering from neurodegenerative diseases, including Alzheimer's disease, mild cognitive impairment, amyotrophic lateral sclerosis, Parkinson's disease or Huntington's disease, and also including neurodegenerative diseases such as Alzheimer's disease, mild cognitive impairment, amyotrophic lateral sclerosis, Parkinson's disease or Huntington's disease. The TF expression variation coefficient is obviously lower than that of a common exosome marker, and the expression level of TF has stronger correlation with the total protein amount of the exosome and is not influenced by external factors such as cell inflammation stress. The TF as the internal reference protein has more excellent performance, can help to more accurately reflect the total loading amount of the sample, and provides a more stable and reliable standardized tool for the field of exosome research.

The invention relates to the technical field of cell culture media, in particular to a sheep testis somatic cell culture medium and a culture method.The sheep testis somatic cell culture medium is prepared from, by weight, 10-15 parts of MEMS alpha, 4-7 parts of KSR, 2-4 parts of sodium chloride, 3-5 parts of recombinant human transferrin, 1-3 parts of ferric citrate, 2-4 parts of L-glutamine, 5-10 parts of water and 4-7 parts of hydroxyapatite. According to the cell culture medium, MEM and KSR are adopted as matrix raw materials, sodium chloride and other auxiliaries are added, the hydroxyapatite agent is matched, the performance change of the cell culture medium on the survival rate and the survival amount of testicular somatic cells is enhanced, meanwhile, hydroxyapatite serves as a matrix of the hydroxyapatite agent, the performance of the cell culture medium is improved through a specific modification solution, and the cell culture medium is prepared. And the function of hydroxyapatite blended kaolin in the system is optimized, so that the functional effect of the cell culture medium is enhanced, and the performance of the product is further improved.

The invention relates to an adipose-derived stem cell culture medium containing wormwood extract and a preparation method of the adipose-derived stem cell culture medium, and belongs to the technical field of biology. The adipose-derived stem cell culture medium contains a DMEM (Dulbecco Modified Eagle Medium) basal culture medium, and also contains the following components: 5 to 7 [mu] g / mL of wormwood extract, 1 to 1.5 [mu] g / mL of phosphatidylserine, 0.02 to 0.03 [mu] g / mL of D-calcium pantothenate, 1.2 to 1.5 [mu] g / mL of sialic acid, 0.06 to 0.08 [mu] g / mL of eucommia male flower peptide, 0.3 to 0.35 [mu] g / mL of transferrin and 0.3 to 0.4 mg / mL of bovine serum albumin. The prepared adipose-derived stem cell culture medium has a good promoting effect on proliferation of adipose-derived stem cells from rabbits and mice, the added wormwood polysaccharide extract has a good bacteriostatic effect, and the safety and stability of the culture medium can be effectively maintained.

The application discloses a kind of R8 and Tf co-modified mebendazole liposome targeted preparation and its preparation method, application, belong to the field of biological medicine. In view of the poor solubility of mebendazole in water, difficult to penetrate blood-brain barrier, the problem of insufficient tumor targeting in brain glioma treatment, the application adopts film hydration-step modification process, and octaarginine (R8) and targeting ligand transferrin (Tf) are simultaneously modified on the surface of mebendazole-loaded liposome, and the preparation prepared by the application has an encapsulation efficiency of not less than 95%. The preparation can break through blood-brain barrier and precisely enrich in brain glioma lesions, and the in-vivo tumor inhibition rate is more than 90%, with low toxicity and side effects. The preparation can be used for preparing brain glioma therapeutic drugs, and provides a new drug for targeted treatment of brain glioma.

Disclosed herein are engineered nucleases that bind and cleave a recognition sequence within intron 1 of a transferrin gene, and methods of using such engineered nucleases to produce a genetically-modified eukaryotic cell comprising a modified transferrin gene. Further provided are pharmaceutical compositions and methods for treatment of a variety of conditions through expression of a polypeptide of interest encoded by an exogenous nucleic acid molecule inserted in intron 1 of a transferrin gene and expressed under the control of the endogenous transferrin promoter.

The invention provides an induced pluripotent stem cell culture medium and a culture method of induced pluripotent stem cells. The induced pluripotent stem cell culture medium comprises a basic culture medium, a composite additive A and a composite additive B, the composite additive A comprises 5-30 [mu] g / mL of recombinant human insulin growth factors, 1-5 ng / mL of recombinant human transforming growth factors, 50-200 ng / mL of recombinant human basic fibroblast growth factors, 7-20 [mu] g / mL of sodium selenite, 0.5-2 mg / mL of albumin, 50-70 [mu] g / mL of ascorbic acid and 5-20 [mu] g / mL of transferrin; the composite additive B is prepared from 50 to 300mg / mL of a Stock B solution, 0.5 to 5 percent (v / v) of a non-essential amino acid solution, 50 to 200ng / mL of DL-piperidinecarboxylic acid, 50 to 200mu g / mL of gamma-aminobutyric acid, 0.1 to 1 percent (v / v) of a lipid concentrated solution, 0.5 to 5 percent (v / v) of an L-alanyl-L-glutamine solution and 0.1 to 0.1 percent of mercaptoethanol. The culture medium for the induced pluripotent stem cells provided by the invention can ensure the self-renewal capability of the iPSCs in a long-term passage process; and the genetic stability of the iPSCs cells is optimized according to the genome integrity and pluripotent state.

The present invention relates to an antitumor adenovirus capable of evading the in vivo immune system. According to the present invention, the adenovirus comprising a nucleic acid coding for a transferrin-binding domain of the present invention exhibits notably increased effects of infecting and killing tumor cells, exhibits increased binding to transferrin, thereby evading an in vivo immune response and thus increasing the plasma half-life, has a systemic therapeutic effect by being specifically delivered to cancer cells, is capable of being topically delivered, and has excellent selectivity, thereby exhibiting the effect of notable antitumor efficacy, and thus may be usefully employed as an anticancer composition or an anticancer adjuvant for various carcinomas.

Antibodies can treat neurodegenerative disorders (NDs) caused by misfolded proteins. Still, the blood-brain-barrier (BBB) resists their entry, a barrier that can be reduced by scFv instead of whole antibody and conjugating it with transferrin protein to induce transcytosis. While the scFv binds variable heavy and light chains with a non-cleavable linker, a cleavable linker between the scFv and transferrin protein can reduce exocytosis, enhancing the activity of the scFv.

Provided are transferrin receptor binding VHH domains as well as delivery complexes and methods for targeting cells of interest for delivery of an active cargo such as an oligomeric agent. In particular, the delivery complex may comprise modified Fc domain and a transferrin receptor binding VHH domain.

The invention belongs to the technical field of biology, and discloses a serum-free culture medium and application thereof in obtaining nerve cells through embryo fibroblast transdifferentiation. The serum-free culture medium consists of a basic culture medium, a protein additive and cell culture auxiliary components, the protein additive consists of transferrin, a leukemia inhibition factor and a basic fibroblast growth factor; the final concentration of the transferrin is 5 ng / mL-30 ng / mL, the final concentration of the leukemia inhibition factor is 50 ng / mL-150 ng / mL, and the final concentration of the basic fibroblast growth factor is 10 ng / mL-25 ng / mL. The serum-free culture medium can promote the conversion speed of transdifferentiation of fibroblasts into nerve cells, can significantly improve the transdifferentiation efficiency, and can improve the cell survival rate of transdifferentiation of mouse embryo fibroblasts into nerve cells.

The invention provides a culture method of human umbilical cord mesenchymal stem cells, and belongs to the technical field of cell culture. The method comprises the following steps: firstly, constructing an activation culture medium aiming at an initial culture stage and an amplification culture medium aiming at an amplification culture stage; wherein the activation culture medium takes alpha-MEM containing 10% of fetal calf serum as a basic culture medium, and alpha-tocopherol, nicotinic acid and gypenoside are added as additives, so that the cell amplification efficiency in the initial stage can be fully improved; a serum-free scheme is adopted for an amplification culture medium, RPMI-1640 is taken as a basic culture medium, and glutamine, rhG-CSF, bFGF, VEGF, transferrin, vitamin C, 2-mercaptoethanol, zoledronic acid, lenalidomide, ethanolamine and sodium selenite with specific amounts are added, so that the cell amplification speed in an amplification culture stage is remarkably increased. On the basis, a three-stage amplification process is adopted, and large-scale culture of the umbilical cord mesenchymal stem cells is realized. The method is short in culture time and high in cell quantity accumulation speed, and the culture efficiency is remarkably improved.

A culture medium for immune cells and use thereof. The culture medium comprises: 200-1000 U / mL IL-2, 10-100 ng / mL IL-4, 10-100 ng / mL IL-15, 2-20 ng / mL anti-CD3 monoclonal antibody, 2-20 ng / mL anti-CD28 monoclonal antibody, 0.5-20 μM GSK-3β inhibitor, 0.1-10 μM Wnt / β-catenin signaling pathway agonist, 1-10 μg / mL transferrin, 1-10 μg / mL ethanolamine, 1-10 μg / mL water-soluble cholesterol, 1-10 μg / mL insulin, 1-10 mM L-alanyl-L-glutamine, 1-10 μg / mL L-ascorbic acid, 1-10 μg / mL taurine, 0.1-1 μg / mL putrescine, and an RPMI 1640 medium. The culture medium can effectively keep the survival rate of cells, efficiently induce the proliferation of memory T cells, and amplify the number of memory T cells. The culture medium has the advantages of low cell culture cost and good cell quality.

The invention provides a biotinylated Angiopep-2 modified targeting nano material as well as a preparation method and application of the biotinylated Angiopep-2 modified targeting nano material. According to the preparation method of the biotinylated Angiopep-2 modified targeting nano-material, Angiopep-2 is directionally anchored through a high-affinity biotin-streptavidin system, ligand conformation inactivation caused by chemical coupling is avoided, targeting activity is maintained, the used Angiopep-2 can help nanoparticles to be specifically recognized and combined and mediate the nanoparticles to cross a blood brain barrier to enter the brain, and the targeting nano-material can be used for preparing the targeted nano-material. The drug concentration in brain tissues is increased, so that the cure rate of brain diseases is improved, the brain targeting efficiency is far higher than that of transferrin and lactoferrin, and the application value is great; by entrapping curcumin and quercetin, a hydrophilic shell is provided, so that the water solubility of curcumin and quercetin is improved, and the bioavailability of curcumin and quercetin is improved.

The invention discloses a culture medium for in-vitro adherent culture of primary tumor cells and application, and relates to the technical field of tumor cell culture. Comprising a basic culture medium, fetal calf serum FBS with the concentration of 5 ng / mL-15 ng / mL, an epidermal growth factor EGF with the concentration of 1 ng / mL-10 ng / mL, a basic fibroblast growth factor bFGF with the concentration of 0.5 ng / mL-5 ng / mL, an insulin-transferrin-selenium ITS supplement with the final volume concentration of 0.1%-1%, L-glutamine with the final volume concentration of 0.5 mM-2 mM, ascorbic acid with the final volume concentration of 0.1%-0.5% and non-essential amino acid with the final volume concentration of 1%-5%. The kit comprises 10 [mu] g / mL-50 [mu] g / mL of heparin and 1 [mu] g / mL-5 [mu] g / mL of an Rho kinase inhibitor. The culture medium overcomes the defects that in the prior art, when primary tumor cells are subjected to adherent culture, apoptosis or dedifferentiation is prone to occurring, in-vivo tumor characteristics are difficult to simulate, the tumor growth speed is low, the proliferation effect is poor, and the adherent rate is low.

The invention belongs to the technical field of cell therapy, and relates to a serum-free and heterologous component-free NK cell culture medium as well as a preparation method and application thereof. The culture medium comprises a classical culture medium, a proliferation promoting component, vitamins, trace elements, lipids, transferrin, an energy substance and beta-mercaptoethanol, the proliferation promoting component is prepared from human albumin, resveratrol, insulin and hydroxyproline. On the basis of a serum-free and heterologous component-free NK cell culture medium, IL-12, rapamycin, polyinosinic-polycytidylic acid and human AB plasma are additionally added to obtain an activated culture medium, the activated culture medium and an amplification culture medium are jointly used, the culture effect is better, and amplification components IL-2 and human AB plasma are additionally added to obtain an amplification culture medium. Through precise component design, the NK cell in-vitro proliferation efficiency, motility, purity and killing ability are remarkably improved, and key problems existing in a traditional culture medium are efficiently solved.

The application discloses a kit for detecting transferrin and application thereof, and belongs to the technical field of transferrin content detection. The kit comprises a PBA-SiO2@Fe3O4 magnetic affinity probe and an Ab-Au, Pt, Cu@PDA nanoscale enzyme immune probe. When the transferrin exists, the PBA-SiO2@Fe3O4 magnetic affinity probe and the Ab-Au, Pt, Cu@PDA nanoscale enzyme immune probe are combined with the transferrin to form a sandwich structure similar to a sandwich. The catalytic performance of the Ab-Au, Pt, Cu@PDA nanoscale enzyme and the Raman enhancement activity of Au, Pt and Cu metal are utilized to make the substrate produce an oxidation product 3,3',5,5'-tetramethyl benzidine diimine with color developing capability, a novel transferrin multi-mode sensing platform with rapidness, sensitivity, selectivity and high reliability is established, and the visual sensing analysis can realize rapid semi-quantitative on-site detection of transferrin in samples. The method can be applied to on-site rapid detection of low-concentration transferrin in sweat, blood, urine and other samples.

The invention discloses an induction culture medium with definite components and application of the induction culture medium in preparation of a hepatic fibrosis treatment medium, and belongs to the technical field of hepatic cell culture and artificial liver. The induction culture medium is composed of a serum-free basic culture medium, primary bile acid and secondary bile acid, the serum-free basic culture medium is composed of a William's E culture medium, penicillin, streptomycin, L-glutamine, insulin, transferrin, sodium selenite and dexamethasone, the primary bile acid is an FXR signal channel stimulant, and the secondary bile acid is a TGR signal channel stimulant. The hepatic fibrosis treatment medium prepared by utilizing the induction culture medium has a remarkable hepatic fibrosis relieving effect, and has the advantages of definite production materials, controllable process, high product stability and the like.

A hematopoietic stem cell (HSC) proliferation medium includes a basal medium, 0.1 ng / mL to 200.0 ng / mL of thrombopoietin (TPO), 0.1 ng / mL to 200.0 ng / mL of stem cell factor (SCF), 0.1 ng / mL to 20.0 ng / mL of interleukin-3 (IL-3), 0.1 ng / mL to 100.0 ng / mL of interleukin-6 (IL-6), 0.1 ng / mL to 200.0 ng / mL of Flt-3 ligand (FL), 0.1 μg / mL to 20.0 μg / mL of insulin, 0.1 μg / mL to 120.0 μg / mL of transferrin, 0.1 g / L to 3.0 g / L of human serum albumin (HSA), 0.1 μM to 4.0 μM of pyrimido-indole derivative UM729, and 0.1 μM to 2.0 μM of aryl hydrocarbon receptor (AhR) antagonist StemRegenin 1 (SR1). Methods for in vitro proliferation of hematopoietic stem cells (HSCs) and in vitro production of hematopoietic progenitor cells (HPCs) using the HSC proliferation medium are also provided.

Disclosed are novel vaccine compositions comprising modulated Transferrin Binding Protein B (TbpB) polypeptides or immunogenic portions thereof. Notably, the TbpB polypeptides are modulated with respect to domains known as loop domains located in the C-terminal lobe of TbpB polypeptides. The vaccine compositions may be used to ameliorate, treat or prevent pathogenic infections in humans caused by pathogenic H. influenzae strains. Related methods and uses are also disclosed.

The present invention provides a primary cell culture medium for culturing primary lung cancer epithelial cells, comprising an MST1 / 2 kinase inhibitor, a ROCK kinase inhibitor, a fibroblast growth factor, at least one additive selected from the group consisting of a B27 additive and an N2 additive, epidermal growth factor, transferrin, gastrin, and a TGFβ type I receptor inhibitor. The present invention also relates to a culture method using the primary cell culture medium and its use in drug efficacy evaluation and screening. The culture method uses the primary cell culture medium to culture primary lung cancer cells on a culture vessel coated with extracellular matrix gel, resulting in rapid proliferation of the primary lung cancer cells. The cell model obtained using the primary cell culture medium and primary cell culture method of the present invention can be used for drug efficacy evaluation and screening.

The invention discloses a skincare stem cell exosome separation and extraction method, and relates to the technical field of biological medicine, the skincare stem cell exosome separation and extraction method comprises the following steps: S1, culture of mixed stem cells: unfreezing urine-derived stem cells and pilose antler stem cells, resuspending in an improved DMEM culture medium, and carrying out mixed culture to prepare a mixed culture solution; epidermal growth factors, cholera toxin, penicillin, streptomycin, transferrin, IL-2 and IL-15 are added into the improved DMEM culture medium; s2, pretreatment of the mixed culture solution: performing centrifugal treatment on the mixed culture solution, and separating to obtain a precipitate and a supernatant, the precipitate being mixed stem cells. The urine-derived stem cells and the pilose antler stem cells are subjected to mixed culture and matched with the improved culture medium containing IL-2, IL-15 and other components, so that the yield and biological activity of the exosome are remarkably improved, high purity and low impurity protein residue of the exosome are achieved by means of a complete purification process of multi-step centrifugation, ultrafiltration concentration and filter membrane filtration, and the yield of the exosome is improved. The skin allergy stimulation risk is effectively reduced.

The invention discloses a preparation method and application of autonomic ganglion organoid, and relates to the technical field of biology. The method comprises the following steps: 1) providing neural crest stem cells; 2) differentiating the neural crest stem cells into autonomic ganglion-like organs through the combined action of added external factors and internal factors; wherein the external factors comprise a bFGF (basic fibroblast growth factor), an IGF-1 (insulin-like growth factor), a BDNF (brain-derived neurotrophic factor), a GDNF (brain-derived neurotrophic factor), an NGF (nerve growth factor), an NT-3, an EGF (epidermal growth factor), Forskolin, a DAPT (diaminopropyltriethoxysilane), an insulin, a Transferrin, an Activin A and a BMP4 (bone morphogenetic protein 4); the addition of the intrinsic factors is that the following four transcription factors, namely Ascl1, Hand2, Phox2a and Phox2b, are overexpressed in the neural crest stem cells at the same time. The organoid prepared by the scheme contains various neurons similar to the autonomic ganglion in vivo, Schwann glial cells and satellite glial cells. The prepared autonomic ganglion organoid can be applied to research on development regulation of autonomic ganglions and occurrence and development mechanisms of various human diseases, development of therapeutic drugs and the like.

The invention provides a serum-free medium for maintaining dryness of mesenchymal stem cells as well as a preparation method and application of the serum-free medium, and belongs to the technical field of biological medicines. The serum-free culture medium comprises a basic culture medium and an exogenous additive component, the basic culture medium is a DMEM / F12 culture medium, and the exogenous additive components comprise bFGF, EGF, PDGF-BB, recombinant human fibronectin, laminin, insulin, transferrin, sodium selenate, beta-mercaptoethanol and ascorbic acid. The serum-free culture medium provided by the invention is a special culture medium for the mesenchymal stem cells, which is completely free of animal-derived components, is suitable for large-scale amplification of clinical-grade cell treatment products, is relatively low in preparation cost and high in stem differentiation capability, and can remarkably improve the proliferation efficiency of the cultured mesenchymal stem cells.

The application discloses a serum-free adipogenic differentiation method of pig fat precursor cells, relates to the technical field of mammal cell differentiation, and uses an induction differentiation culture medium and a maintenance differentiation culture medium in succession in the adipogenic differentiation process. The induction differentiation culture medium is composed of a basic culture medium and a cell culture auxiliary factor. The cell culture auxiliary factor is composed of 66nmol / L insulin, 33μmol / L biotin, 17μmol / L pantothenic acid, 0.1μmol / L dexamethasone, 0.25mmol / L IBMX, 1μmol / L rosiglitazone, 150μmol / L sodium oleate and 10μg / mL transferrin. The maintenance differentiation culture medium removes the IBMX and rosiglitazone on the basis of the induction differentiation culture medium. The serum-free adipogenic differentiation method of pig fat precursor cells has good differentiation effect and high repeatability.

Antibodies can treat neurodegenerative disorders (NDs) caused by misfolded proteins. Still, the blood-brain-barrier (BBB) resists their entry, a barrier that can be reduced by scFv instead of whole antibody and conjugating it with transferrin protein to induce transcytosis. While the scFv binds variable heavy and light chains with a non-cleavable linker, a cleavable linker between the scFv and transferrin protein can reduce exocytosis, enhancing the activity of the scFv.

This invention belongs to the field of immune cell culture technology, specifically relating to an in vitro expansion culture medium for NK cells and its culture method. It consists of a basal culture medium and auxiliary additives. The basal culture medium is X-VIVO 15 medium; the auxiliary additives include the following components: human serum albumin (HSA), IL-12, IL-18, IL-15, rapamycin, nicotinamide, gelatin-IL-15 microspheres, alginate-IL-21 microspheres, recombinant human insulin, transferrin, glutamine, β-mercaptoethanol, a mixture of non-essential amino acids and lipids. This invention achieves time-dependent synergistic IL-15 sustained-release / IL-21 pulsed release and metabolic synergistic NAD+ release. + Mitochondrial / lipid membrane homeostasis enables efficient amplification and long-lasting cytotoxicity.

The present invention provides a culture medium and a culture method for primary mesothelial cells. The culture medium comprises a basal culture medium and one or more additives selected from the group consisting of a Rho proteinase inhibitor, epidermal growth factor, hydrocortisone, a glutamine additive, amphiregulin, pancreatic transferrin, and fetal bovine serum. The use of the culture medium for primary mesothelial cells of the present invention creates a more suitable living environment for the growth and reproduction of primary mesothelial cells, thereby significantly improving the culture success rate and expansion rate of primary mesothelial cells.