29 results about "Breast epithelial cell" patented technology

A three-dimensional mammary gland model comprising a three-dimensional tissue comprising cells and fragmented extracellular matrix components, the cells comprising mature adipocytes and mammary gland epithelial cells.

The present invention discloses a targeted knockout bovine FGFR1 The application of the sgRNA of the gene in constructing a bovine mammary epithelial cell line, wherein the sgRNA nucleotide sequence is shown in SEQ ID NO: 1. The mammary epithelial cell line obtained by constructing the sgRNA of the present invention through CRISPR / Cas9 technology, FGFR1 Protein expression was significantly downregulated, and knockout FGFR1 The gene expression had no significant effect on the viability and proliferation of mammary epithelial cells. PCNA 、 Cyclin D1 、 Cyclin E1 There was no significant change in the mRNA expression of the genes related to milk protein synthesis (such as CSN1S1 、 CSN2 、 CSN3 、 EFL5 ), lactose synthesis-related genes ( GLUT1 and HKⅡ ) and milk fat synthesis-related genes ( SCD‑1 ) were significantly decreased, providing a basis for in-depth analysis FGFR1 It provides key tools for the regulatory mechanisms of mammary epithelial cell proliferation, lactation synthesis and other biological behaviors.

The present disclosure pertains to a method for producing a three-dimensional mammary gland model that comprises a three-dimensional tissue body including mature adipocytes and mammary epithelial cells, the method comprising: a step for gelling a mixture that contains a fragmented extracellular matrix component, an alginate gel precursor, and cells that include adipose-derived stem cells and mammary epithelial cells; and a step for subjecting the gelled mixture to differentiation induction culture to thereby differentiate at least a portion of the adipose-derived stem cells into the mature adipocytes to form the three-dimensional tissue body.

The invention provides a method for inducing transdifferentiation of somatic cells into mammary epithelial cells by using an exosome for inducing mammary epithelial cells. The method is efficient, safe and high in specificity. The somatic cells are induced to be transdifferentiated into the mammary epithelial cells by inducing the mammary epithelial cell exosome, so that a new cell source and a new technical means are provided for repairing and regenerating mammary tissues and treating related diseases.

A method for the precise cellular destruction of an oncogenic non-coding RNA with a RNA-binding small molecule conjugated with bleomycin A5 is described. The method affords reversal of phenotype. Bleomycin A5 was coupled to an RNA-binding molecule that selectively binds the microRNA-96 hairpin precursor (pri-miR-96). By coupling of bleomycin A5's free amine to the RNA-binding molecule, its affinity for binding to pri-miR-96 is >100-fold stronger than to DNA. The conjugate compound selectively cleaves pri-miR-96 in triple negative breast cancer (TNBC) cells. Selective cleavage of pri-miR-96 enhances expression of FOXO1 protein, a pro-apoptotic transcription factor that miR-96 silences, and triggers apoptosis in TNBC cells. No effects were observed in healthy breast epithelial cells. This method provides programmable control for targeting RNA through the selection of an RNA-binding molecule / bleomycin A5 conjugate and provides a facile method of mapping the cellular binding sites of an RNA-binding molecule.

The invention belongs to the technical field of biology, and particularly relates to an SgRNA expression vector as well as a construction method and application thereof. The SgRNA expression vector is formed by connecting a pGL3-U6-sgRNA-PGK-puromycin vector with a gRNA1 (guide ribonucleic acid 1), or is formed by connecting a pGL3-U6-sgRNA-PGK-puromycin vector with a gRNA2 (guide ribonucleic acid 2), and the SgRNA expression vector is formed by connecting a pGL3-U6-sgRNA-PGK-puromycin vector with a gRNA2 (guide ribonucleic acid 2); the sequence of the gRNA1 is as shown in SEQ ID NO. 1, and the sequence of the gRNA2 is as shown in SEQ ID NO. 2. Respectively mixing the mRNA of the ABE7.10 and the transcription mRNA of the gRNA1, and mixing the mRNA of the CGBE and the transcription mRNA of the gRNA2, and injecting the mixture into a fertilized egg or co-injecting the mixture into the fertilized egg to obtain the gene editing cloned cattle. Experimental results show that the expression quantity of the proinflammatory factors of the mammary epithelial cells separated from the gene-edited cloned cattle is remarkably reduced.

The invention relates to the technical field of biological medicines, in particular to mRNA (messenger Ribonucleic Acid) for coding a single-chain antibody resisting staphylococcus aureus and application of the mRNA. On the basis of a specific scFv with high affinity to staphylococcus aureus lysate, cell wall protein and toxin, the mRNA is optimized by a gene sequence and is matched with a functional element suitable for in-vivo expression of the mRNA, so that stable and efficient expression of the mRNA in a target tissue is ensured. The mRNA is further prepared into an mRNA-LNP preparation, and the preparation is beneficial to delivery of the mRNA to local tissues of the mammary gland of the dairy cow, expresses scFv protein with a specific antibacterial function in epithelial cells of the mammary gland, can be used for replacing traditional antibiotics to treat the mastitis of the dairy cow caused by staphylococcus aureus, and has an outstanding treatment effect.

The present application belongs to the technical field of natural polymer materials, and particularly relates to a purified polysaccharide of Rhizoma Polygonati, a preparation method thereof and application of the purified polysaccharide in preparation of a medicine for relieving mammary gland hyperplasia. The purified polysaccharide of Rhizoma Polygonati comprises xylose, mannose, glucose and galactose, and the molar ratio of the xylose, mannose, glucose and galactose is 6.45:30.26:7.43:55.87. The molecular weight distribution of the purified polysaccharide of Rhizoma Polygonati is within 2000-30000 Da, and the molecular weight is concentrated at 22877 Da. The purified polysaccharide of Rhizoma Polygonati can reduce the toxic damage of estrogen and progesterone imbalance to HC11 mammary epithelial cells, and a low concentration of the purified polysaccharide of Rhizoma Polygonati can produce a protective effect on the cells. The purified polysaccharide of Rhizoma Polygonati is verified to have a significant inhibitory effect on mammary gland hyperplasia of mice through a mammary gland hyperplasia model intervention experiment, and therefore can be used for developing a medicine for treating mammary gland hyperplasia.

A three-dimensional mammary gland model includes a three-dimensional tissue containing cells and fragmented extracellular matrix components, and the cells include mature adipocytes and mammary epithelial cells.

A culture medium for culturing mammary epithelial cells, which medium contains β-estradiol, insulin-like growth factor 1, basic fibroblast growth factor, tumor necrosis factor-α, and optionally fibroblast growth factor 10. A method for culturing mammary epithelial cells using the culture medium, and a method and the use of cells obtained using the culture method in evaluating or screening the efficacy of a drug.

The application discloses a method for improving the passaging times of a mammary epithelial cell line and application thereof, and belongs to the technical field of mammary epithelial cell lines. The application first finds that the exogenous fragment HS4-SAR fragment in the attachment area of the insulator-scaffold is connected with a p-Ef1a-SV40LT-Puro vector or a pEF1a-eGFP vector, and the transfection of commercial immortalized cell lines and primary cell lines can significantly improve the passaging times of the mammary epithelial cell lines, the cell morphology is still normal after 120 generations, the malignant transformation does not occur, the gene expression level does not decrease, and the immortalization is realized; the established goat mammary epithelial cell line still has the ability of synthesizing and secreting proteins; the method for improving the passaging times of the mammary epithelial cell line is simple and easy to operate, has a significant effect on the immortalized cell lines and the primary cell lines, and is expected to improve the stable passaging times of various mammary epithelial cell lines, and reduce the cell cost of related research.

The invention relates to the technical field of molecular biology, in particular to an MUC15 gene segment, expression vector construction of the MUC15 gene segment and an expression vector transfection method of the MUC15 gene segment, and the nucleotide sequence of the MUC15 gene segment influencing buffalo mammary epithelial cell triglyceride and fatty acid synthesis is shown as SEQ No.1. The MUC15 gene segment is obtained by amplifying a primer pair, and the primer pair is used for amplifying the buffalo mammary epithelial cell triglyceride and fatty acid synthesis. The primer pair is specifically as follows: F: 5 '-AGTCCGGACTCAGATCTCGAGATGTTGACTCAGCCAAAATTCTG-3', F: 5 '- And R: 5 '-AATTCGAAGCTTGAGCTCGAGCTTACACGAAGTGGAGG-3'. Compared with the prior art, the buffalo mammary epithelial cell MUC15 gene fragment influencing the synthesis of triglyceride and fatty acid in the buffalo mammary epithelial cell is obtained, and a pEGFP-C1-MUC15 eukaryotic expression vector is constructed and transfected to the buffalo mammary epithelial cell, so that the synthesis of triglyceride by the buffalo mammary epithelial cell is promoted, and the milk fat content in buffalo milk is increased.

A method of producing mammalian breast milk exosomes, such as human breast milk exosomes, comprising generating lactating cells derived from mammalian breast epithelial cells, such as human breast epithelial cells, expressing mammalian milk-like products, such as human milk-like products, from the lactating cells, and purifying the exosomes from the mammalian milk-like products.

The invention provides application of a ceramide transporter mutant in promoting in-vitro cell synthesis of sphingomyelin, and relates to the technical field of biosynthesis, the amino acid sequence of the ceramide transporter mutant is as shown in SEQ ID NO.1, and cells comprise at least one of mammary epithelial cells, stem cell-like cells or embryonic kidney cells. Experiments prove that the ceramide transporter mutant can promote mammary epithelial cells or stem cell-like cells to synthesize sphingomyelin in vitro, the yield of sphingomyelin synthesized by the cells in vitro is high, and the ceramide transporter mutant is suitable for large-scale industrial application. The technical problems that in the prior art, the yield of sphingomyelin is low, the process cost is high, and industrial large-scale production is limited are solved.

Provided herein are cell constructs for producing a cultured milk product from mammary epithelial cells (MECs). In some embodiments, the cell constructs comprise a scaffold, a culture medium in fluidic contact with the scaffold, and mammary cells coupled to the scaffold. In some embodiments, one or more features and / or properties of the scaffold are specified so as to mimic a basement membrane to help to induce the secretory phenotype of mammary epithelial cells in vitro.

The present disclosure relates to the technical field of biomedicine. Provided in the present disclosure are an induction medium and method for promoting mammary epithelial cells to secrete milk-like exosomes. Regarding the induction medium, estrogen and glucose are added to an MEpiCM medium, wherein the molar concentration of the estrogen is 50 to 200 μM, and the molar concentration of the glucose is 10 to 30 mM; and the pH of the induction medium is 6.4 to 7.1. An acidic microenvironment is formed by means of adjusting the pH, which increases the permeability of cell membranes and activates intracellular signaling pathways. In addition, 50 to 200 μM of estrogen and 10 to 30 mM of glucose are added, which interact with the acidic microenvironment formed by means of pH adjustment, thereby making the estrogen and glucose to regulate mammary epithelial cells and inducing the mammary epithelial cells to secrete exosomes.

The invention relates to the technical field of molecular biology, in particular to an ACSL3 gene segment, an expression vector construction method thereof and an expression vector transfection method thereof, the nucleotide sequence of the ACSL3 gene segment influencing buffalo mammary epithelial cell triglyceride and fatty acid synthesis is shown as SEQ No.1, the ACSL3 gene segment is obtained by amplifying a primer pair, and the primer pair has a nucleotide sequence shown as SEQ No.2. The primer pair is specifically as follows: F: 5 '-AGTCCGGACTCAGATCTCGAGATGAATAACCAGGGTCTACAA-3', F: 5 '-AG: 5'-AG: 5 '-AG: 5'-AG: 5 '-AG: 5'-AG: 5 ' And R: 5 '-AATTCGAAGCTTGAGCTATGAGCTATTTTCTTCCATATTCGCTCA-3'. Compared with the prior art, the ACSL3 gene fragment influencing the synthesis of the buffalo mammary epithelial cell triglyceride and fatty acid is obtained, and a pEGFP-C1-ACSL3 eukaryotic expression vector is constructed and transfected to the buffalo mammary epithelial cell, so that the synthesis of the triglyceride by the buffalo mammary epithelial cell is promoted, and the milk fat content in buffalo milk is increased.

The invention belongs to the technical field of biology, and particularly relates to a method for preparing human lysozyme exosome fusion protein based on immortalized bovine mammary epithelial cells and application of the human lysozyme exosome fusion protein. The method comprises the following steps: acquiring and purifying primary bovine mammary epithelial cells; transfecting an hTERT gene to construct an immortalized cell line; constructing an expression plasmid containing the fusion gene of the CD47 ankyrin and the human lysozyme; transfecting plasmids into immortalized cells, and screening positive clones; and finally, separating the human lysozyme exosome fusion protein from the cell culture supernatant by using a Ni-NTA chromatographic column to obtain the human lysozyme exosome fusion protein. The fusion protein has lysozyme activity, can effectively dissolve bacteria, has antibacterial and anti-inflammatory functions, is high in stability and low in antigenicity and is suitable for preparing antibacterial or anti-inflammatory drugs, immortalized cells are used as a bioreactor, efficient and continuous secretion of human lysozyme is achieved, and the human lysozyme can be used for preparing antibacterial or anti-inflammatory drugs. The problems of low yield, poor activity and insufficient stability in the traditional method are solved.

The invention provides application of myricetin-3-o-galactoside in induction of casein expression in mammary epithelial cells, and belongs to the technical field of cell culture. The invention provides an application of myricetin-3-o-galactoside in at least one of the following aspects: promoting cell growth, inducing casein expression in cells and constructing a casein expression dominant cell model. The myricetin-3-o-galactoside can be used for inducing the expression of casein in cells and promoting the growth of the cells. The embodiment of the invention shows that after myricetin-3-o-galactoside is added into the culture medium, the casein expression quantity in mammary epithelial cells is improved by 5.1 times, and the cell growth rate is also obviously improved. In addition, the method is high in repeatability and stable in induction effect, and a new path is provided for improving the yield and quality of dairy products.

A culture medium for culturing mammary epithelial cells, comprising β-estradiol, insulin-like growth factor 1, basic fibroblast growth factor, tumor necrosis factor-α, and optionally fibroblast growth factor 10. A method for culturing mammary epithelial cells using said culture medium, as well as a method and the use of cells obtained using said culture method in evaluating the efficacy of drugs or screening drugs.

The invention provides isolated human breast milk exosomes, and compositions thereof. The present invention also provides in vitro methods for producing said exosomes comprising generating lactocytes derived from mammalian mammary epithelial cells, for example human mammary epithelial cells, expressing the mammalian milk like product, for example the human milk like product from lactocytes, and purifying the exosomes from the mammalian milk like product. The invention also provides therapeutic uses of the isolated human breast milk exosomes, and compositions thereof.

The invention provides application of a PPP1R15A gene in regulation and control of oxidative stress of cells, and belongs to the technical field of gene engineering. Experiments show that oxidative stress of bovine mammary epithelial cells can be inhibited by inhibiting expression of the PPP1R15A gene. The invention also provides an application of the siRNA for knocking out or inhibiting expression of the PPP1R15A gene in preparation of a medicine for inhibiting oxidative stress of cells, wherein the sequence of the siRNA is shown as SEQ ID NO.11 and SEQ ID NO.12. The invention also provides an application of the siRNA for knocking out or inhibiting expression of the PPP1R15A gene in preparation of a medicine for inhibiting oxidative stress of the cells. The siRNA provides a new direction for searching a novel anti-oxidative stress treatment strategy.

The invention discloses application of salidroside in preparation of a medicine for relieving hypoxia injury of mammary epithelial cells of dairy cows, and relates to the field of biological medicine, and the salidroside (Saldroside, Sal) can relieve hypoxia stress in the mammary epithelial cells (BMECs) by inhibiting mitochondrial autophagy mediated by BNIP3 / NIX and regulating mitochondrial metabolism. The research can provide a special perspective for the development and utilization of salidroside, and provides a theoretical basis for the development of a new drug for treating the hypoxia injury of the buffalo.

The invention discloses an application of sgRNA for targeted knockout of a bovine FGFR1 gene in construction of a bovine mammary epithelial cell line. The nucleotide sequence of the sgRNA is as shown in SEQ ID NO: 1. According to the mammary epithelial cell line constructed and obtained by utilizing the sgRNA through a CRISPR / Cas9 technology, the FGFR1 protein expression quantity is remarkably reduced, the activity and proliferation condition of mammary epithelial cells are not remarkably influenced after the FGFR1 gene is knocked out, mRNA expression of proliferation key genes PCNA, Cyclin D1 and Cyclin E1 in the cells is not remarkably changed, and the mammary epithelial cell line can be used for preparing a mammary epithelial cell line. However, expression of milk protein synthesis related genes (such as CSN1S1, CSN2, CSN3 and EFL5), lactose synthesis related genes (GLUT1 and HKII) and milk fat synthesis related genes (SCD-1) is remarkably reduced, and a key tool is provided for deep analysis of a regulation mechanism of FGFR1 in biological behaviors such as mammary epithelial cell proliferation and lactation synthesis.

According to the application of the induced mammary epithelial cell exosome in preparation of the medicine for promoting lactation, induced mammary epithelial cells are obtained through a specific induction factor RepSox, the exosome with efficient lactation promoting activity can be secreted, and compared with a traditional lactation promoting method, the application has the advantages of being accurate in action target spot, high in safety and small in side effect.

The invention discloses a cascade nano-enzyme with inflammation targeting ability, which is characterized in that MnO2 is successfully loaded on Ce-MOF-808 / CeO2 through a hydrothermal method, an HA / Ce-MOF-808 / CeO2 / MnO2 nano-enzyme is successfully prepared through an HA coating, the catalase activity of the cerium-based MOF nano-enzyme is enhanced by introducing MnO2, and a cascade nano-platform with ideal enzyme activity is constructed. Meanwhile, hyaluronic acid is coated on the surface of the nano-platform, so that the nano-platform is endowed with the capability of over-expressing a CD44 receptor on the surface of a targeted inflammatory cell, and the nano-platform is used for treating porcine mammary epithelial cell inflammation and has a wide application prospect in the aspect of treating oxidative stress and inflammatory diseases of sows.

Provided is a method of increasing lipid synthesis or accumulation in mammary epithelial cells (MECs) culture and optionally secretion therefrom. The method comprising contacting the MECs with an LXR agonist in the presence of a fatty acid to thereby increase lipid synthesis or accumulation and optionally secretion of lipids. Also provided is a method of producing synthetic fat globules, the method comprising: (a) isolating lipid droplets from cells in culture; (b) encapsulating the droplets with a lipid bilayer, thereby producing synthetic fat globules. Also provided are compositions produced thereby.

The invention belongs to the technical field of biological medicines, and relates to application of exogenous mitochondria in preparation of a medicine for treating colitis, and the exogenous mitochondria is derived from mammary epithelial cells. The exogenous mitochondria derived from mammary epithelial cells is used for treating young individual colitis for the first time, and the symptoms including inhibition of weight loss, improvement of diarrhea and hemafecia, repair of colonic mucosal lesion (such as crypt structure damage and inflammatory cell infiltration) and / or reduction of disease activity index (DAI score) can be remarkably relieved; the problems that in the prior art, mitochondria sources are limited, oral stability is poor, and the curative effect verification of a young model is insufficient are solved, and a new strategy is provided for colitis treatment.

The application provides a feed additive and application thereof. The feed additive comprises the following components in parts by weight: 20-30 parts of flaxseed, 10-20 parts of perilla seed, 1-2 parts of choline, 0.05-0.15 parts of vitamin E, 0.1-0.2 parts of zinc sulfate, 0.01-0.05 parts of nicotinic acid and 48-68 parts of wheat bran. The feed additive and the feed containing the feed additive provided by the application are used for 6-13 month old non-pregnant cows, the mammary gland development of the cows in puberty is intervened, the growth of the mammary gland fat pad in the period is promoted, the growth and development of the lactiferous duct in the gestation period are provided with a material basis, the number of the lactiferous duct and the acinus is increased, and thus the initial number of the mammary epithelial cells in the lactation period is increased, and the peak milk yield of the cows after delivery is improved.