97 results about "Glandular Epithelial Cells" patented technology

The present invention is directed to sensitive and specific methods and kits for the detection of prostate cancer in a patient. The invention uses RT-PCR to detect the expression or change in expression of PSA in epithelial cells enriched from whole blood. The methods and kits on the invention represent a a significant improvement over previous methods of CaP diagnosis. According to one embodiment, prostate epithelial cells are isolated from blood and the RNA subjected to RT-PCR. The resulting cDNA is subjected to traditional PCR amplification with primers able to distinguish between the genomic copy of the gene and the cDNA copy resulting from CaP gene expression. This method provides an assay which is more sensitive, specific and reproducible as compared to conventional methods. Results disclosed herein demonstrate a correlation between the presence of PSA-expressing prostate epithelial cells with cancer recurrence which provides a diagnostic tool for the early detection of prostate disease.

The present invention provides a bioartificial lacrimal gland which contains at least one unit that includes (a) a permeable housing having an interior and an exterior; (b) an outlet connecting the housing interior to the housing exterior; and (c) a population of lacrimal epithelial cells within the housing interior.

The invention provides serum-free culture fluid for culture of salivary gland epidermal cells and salivary gland stem cells of mammals, which is prepared by using MCDB153HAA as basic culture medium and adding amino acid, vitamins, salts, lipid, trace elements, buffer fluid, hormone-like compounds, transgenic metalloprotein, antioxidants, seralbumin, glucide, purine, pyrimidine base substances and pH value indicators. The serum-free culture fluid has the advantages that salivary gland epidermal cells of the mammals can vigorously grow, good cell activity and physiological properties are maintained, in addition, the serum-free culture fluid is also suitable for the salivary gland epidermal cell culture, is particularly suitable for the field of study relevant to biological tissue engineering and belongs to the commercial cell culture fluid.

The invention provides a method for treatment or prophylaxis of benign prostatic hyperplasia by administration of an agent that interferes with energy metabolism, particularly the production of ATP and NADH / NADPH, in prostate epithelial cells.

A reconstituted human breast tumor model is disclosed. The model, which is incorporated into mice, provides actual tumors that arise spontaneously, thereby mimicking naturally occurring breast cancer. The tumors are genetically human, because they arise from human mammary tissues that develop from human mammary epithelial cells implanted into host mice. Prior to implantation, the mammary epithelial cells are genetically modified to contain a recombinant human oncogene and an SV40 early region.

MicroRNA markers of breast cancer stem cells (BCSC) are provided herein. The markers are polynucleotides that are differentially expressed in BCSC as compared to normal counterpart cells. Uses of the markers include use as targets for therapeutic intervention; as targets for drug development, and for diagnostic or prognostic methods relating to breast cancer and BCSC cell populations. BCSCs have the phenotype of having lower expression of certain miRNAs compared to normal breast epithelial cells, or to cancer cells that are not cancer stem cells.

The present invention is directed to the use of antibodies or binding portions thereof, probes, ligands, or other biological agents which either recognize an extracellular domain of prostate specific membrane antigen or bind to and are internalized with prostate specific membrane antigen. These biological agents can be labeled and used for detection of cancerous tissues, particularly cancerous tissues proximate to or containing vascular endothelial cells, which express an extracellular domain of prostate specific membrane antigen. The labeled biological agents can also be used to detect normal, benign hyperplastic, and cancerous prostate epithelial cells or portions thereof. They also can be used alone or bound to a substance effective to ablate or kill such cells as a therapy for prostate or other cancers. Also disclosed are four hybridoma cell lines, each of which produces a monoclonal antibody recognizing extracellular domains of prostate specific membrane antigens of normal, benign hyperplastic, and cancerous prostate epithelial cells or portions thereof.

Described herein are reprogramming techniques allowing for production of mammary-derived iPSCs (“m-iPSCs”). The m-iPSCs described herein exhibit all the hallmarks of stem cell identity including round cluster, bright colony morphology, clonal expansion, and pluripotent marker expression (alkaline phosphatase expression, Oct-4, nanog, etc.) Further refined techniques allow for generation of m-iPSCs under essentially defined conditions.

The invention discloses a primary isolated culture method for dairy cow mammary epithelial cells. The primary isolated culture method comprises steps as follows: mammary tissue of a dairy cow in the lactation period is shorn into tissue blocks, the tissue blocks are digested with collagenase / hyaluronidase digestive juice, a culture dish is inoculated with the digested chylous tissue blocks for culture, cells get free from tissue on peripheries of the tissue blocks after 1-2 days and grow along the wall of the culture dish, fusiform fibroblast is removed with a mechanical scraping method every day during culture, and initial passage is performed when a large quantity of cobblestone-like mammary epithelial cells grow in clusters and in patches and are mutually fused after about 4-5 days. During initial passage, a little fibroblast which is probably mixed is eliminated with trysin with a differential digestive method, and the dairy cow mammary epithelial cells with high purity are obtained. The mammary epithelial cells can emigrate from chylous tissue in a short time; a little mixed fibroblast is removed mainly with a mechanical scraping method, and accordingly, the obtained mammary epithelial cells are seldom damaged and are high in purity.

Prostasin protein has been found to be a useful marker for determination of the invasiveness of and as a means to treat human carcinomas. Using RT-PCR and western blot analyses, prostasin protein and mRNA expression were found in normal human prostate epithelial cells and the human prostate cancer cell line LNCaP, but not in the highly invasive human prostate cancer cell lines DU-145 and PC-3. Imunohistochemistry studies of human prostate cancer specimens revealed a down-regulation of prostasin in high-grade tumors. Using RT-PCR and western blot analyses, prostasin protein and mRNA expression were found in a non-invasive human breast cancer cell line, MCF-7, while invasive human breast cancer cell lines MDA-MB-231 and MDA-MB-435s were found not to express either the prostasin protein or the mRNA. A non-invasive human breast cancer cell line, MDA-MB-453, was shown to express prostasin mRNA but not prostasin protein. Transfection of DU-145 and PC-3 cells with a full-length human prostasin cDNA restored prostasin expression and reduced the in vitro invasiveness by 68% and 42%, respectively. Transfection of MDA-MB-231 and MDA-MB-435s cells with a full-length human prostasin cDNA restored prostasin expression and reduced the in vitro invasiveness by 50% for either cell line. The prostasin gene promoter region was found to be hypermethylated at specific sites in invasive cancer cells.

The invention discloses a method for separating, purifying and culturing a mammary epithelial cell of a ruminant animal, which includes the following steps of: 1) the sampling of a mammary tissue; 2) the preparing of a culture fluid; 3) the culturing of a tissue block for obtaining a primary cultured mammary epithelial cell; 4) the purifying of the mammary epithelial cell for obtaining a purified mammary epithelial cell; and 5) the subculturing of the mammary epithelial cell. The invention simultaneously provides a preservation method for the mammary epithelial cell of the ruminant animal, which includes the following steps of: 1) preparing a freezing protection fluid which comprises the mammary epithelial cell of the ruminant animal; and 2) gradually and slowly reducing the temperature of the freezing protection fluid. The method can be adopted for prolonging the preservation time of the mammary epithelial cell of the ruminant animal.

The present invention provides compositions and methods for the suppression of mammary epithelial cell proliferation. In particular, the present invention provides compositions and methods for IKKα kinase inhibition in breast cancer therapy that is relatively free of toxic side effects.

The invention discloses a method for efficiently separating and purifying mammary epithelial cells. The method is suitable for a plurality of mammals. The mammary tissues of adult mammals, which are in the middle and later periods of gestation or in the lactation period, are collected through aseptic operation. According to the provided method, the separation and purification can be achieved in one step, the whole experiment is simple and efficient; the purification, which comprises repeated differential digestion and differential attachment, in the conventional method is eliminated, the purity of mammary epithelial cells obtained by the provided method can reach 98% or more, the obtained mammary epithelial cells can be continuously cultured in-vitro and passed down for 40 generations, and the lactating function of mammary epithelial cells is not affected. The obtained mammary epithelial cells can be applied to the researches on growth of mammary gland, lactating mechanism, mammary bioreactor, transgenic animals, mammary diseases, and the like.

MicroRNA markers of breast cancer stem cells (BCSC) are provided herein. The markers are polynucleotides that are differentially expressed in BCSC as compared to normal counterpart cells. Uses of the markers include use as targets for therapeutic intervention; as targets for drug development, and for diagnostic or prognostic methods relating to breast cancer and BCSC cell populations. BCSCs have the phenotype of having lower expression of certain miRNAs compared to normal breast epithelial cells, or to cancer cells that are not cancer stem cells.

The invention discloses an application method of astragalus polysaccharide in antagonism of dairy cow mammary epithelial cells apoptosis. The method comprises the following steps: (1) obtaining 2-3 generations of dairy cow mammary epithelial cells; (2) grouping and processing cells; (3) detecting cell survival rate employing a CCK-8 method; (4) detecting a cell cycle employing flow cytometry; (5) detecting the change condition of nucleic acid of apoptotic cells employing a DNA ladder; (6) observing the internal morphological structure change of the cells through a transmission electron microscope; (7) detecting inflammation-associated cytokines content in a cell supernatant employing radioimmunoassay; and (8) detecting inflammation-related enzyme activity in the supernatant employing a biochemical kit method. Through the method disclosed by the invention, the action of the astragalus polysaccharide for antagonism of dairy cow mammary epithelial cells apoptosis is discussed; and the method is efficient, simple, convenient, feasible, and accurate in result.

The invention provides a method for establishing a lactation model of cow mammary gland epithelial cells for researching the milk protein gene expression regulation mechanism and providing favorable conditions for testing a mammary gland expression vector. The testing method comprises the following steps: carrying out primary culture and identification of the cow mammary gland epithelial cells; determining the secretion situation of lactose by using the HPLC method; determining the secretion situation of beta-casein protein by using the HPLC method; and carrying out cryopreservation and recovery on the cells. The method adopts the tissue block method to culturing the cow mammary gland epithelial cells in different periods, compared with other primary culture methods of the mammary gland epithelial cells, such as enzyme digestion and the like, the method does not affect the activity of the mammary gland epithelial cells, the culture cost is very low, and the operation is very simple. The method realizes the continuation of normal culture of a cow mammary gland epithelial cell system, thereby property and effectively preserving a large number of precious cow mammary gland tissue materials. The method provides the important testing materials and a technical platform for researching the lactation mechanism of the cow mammary gland during the different development periods.

The invention discloses a method for building a dairy cattle breast acinus lactation model in vitro, and relates to a method for building a lactation model in vitro. The method solves the problem of low authenticity of in-vivo environment caused by taking a mammary epithelial cell as a research carrier in the existing in-vitro study of the lactation function of the dairy cattle breast. The method comprises the following steps: 1, preparing a tissue culture plate of matrix gel; 2, digesting primary cultured dairy cattle mammary epithelial cell gestated for 4 months by adopting pancreatin and EDTA, and collecting cells after centrifugalization; 3, preparing cell suspension and diluting; and 4, adding the cell suspension into the tissue culture plate of the matrix gel, culturing for 15 days in an incubator to finish the building. The artificial building process of the dairy cattle breast acinus lactation model built by the invention is a simulation process of in-vitro simulation of a galactemia protective screen; and a three-dimensional acinus structure thereof can truly simulate in-vivo situation, has the same lactation function and biological characteristics as the breast acinus in a ruminant, and has high authenticity of in-vivo environment simulation.

The invention discloses a frozen stock solution for storing mammary epithelial cells of a milk goat. The frozen stock solution is prepared by mixing a basic frozen stock solution and insulin-transferrin-sodium selenate in a volume ratio of 100 to (1-2). The basic frozen stock solution is prepared by mixing 10% by volume percent of dimethyl sulfoxide, 30-50% by volume percent of a DMEM / F12 culture medium and 40-60% by volume percent of fetal calf serum. The freeze-saving frozen stock solution disclosed by the invention is few in step, simplified in process and low in cost, the resuscitated mammary epithelial cells are strong in anti-apoptosis performance, oxidization resistance and multiplication capacity, the resuscitation rate of the mammary epithelial cells can be increased, and the vitality of the resuscitated cells reaches up to above 95%, so that the cell growth quality is ensured, the proliferation cycle of cell resuscitation is shortened and the mammary epithelial cells of the milk goat can be saved for a long time.

A cell culture article includes a porous substrate having a plurality of pores and a plurality of interstices in communication with the pores. At least some of the plurality of pores and interstices are sufficiently large for two or more mammary epithelial cells to cluster within the pores or interstices. Non-malignant mammary epithelial cells or breast cancer cells may not attach strongly to the substrate surface, which may encourage cell-cell interaction. In many cases, the article is desirably free of components of unknown origin. The articles may be capable of maintaining culture of malignant and non-malignant mammary epithelial cells and allowing for development of in vivo-like morphologies or characteristics of such cells.

The invention provides a method for constructing a milk cow blood milk barrier three-dimensional model in vitro. The method comprises the steps of cultivation of mammary epithelial cells and fibroblast of a milk cow, cultivation of venous endothelial cells of the milk cow, immunofluorescent identification of the mammary epithelial cells and the venous endothelial cells, cocultivation of Transwell,preparation of a cocultivation medium by taking prolactin, insulin, an epidermal growth factor, cow growth hormone, a vascular endothelial growth factor and a fibroblast growth factor as hormone andcell factors related to growth of the mammary epithelial cells, the fibroblast and the endothelial cells; and finally the epithelial cells, the fibroblast and the endothelial cells are inoculated on atranswell chamber, and accordingly the milk cow blood milk barrier model is successfully constructed. The method can be used for researching a molecular mechanism by which nutrient substances, drugsand toxic substances enter the mammary tissue and the milk in real-time and effectively, not only fills in the gap both domestically and internationally, but also provides a research platform for research on the molecular mechanism by which lipid, vitamins, the drugs and the toxic substances pass through the blood milk barrier.

The invention discloses a cell in-vitro culture method for primary culture of bovine mammary epithelial cells.The cell in-vitro culture method comprises the following steps: pretreating bovine mammary tissues and then standing and carrying out adherent culture for 30 minutes by laterally placing a culture bottle; carrying out isolated culture by using a tissue block recycling method; and finally purifying by using a time-difference trypsin digestion method and a bottle internal digestion method to obtain the bovine mammary epithelial cells.The cell in-vitro culture method for primary culture of the bovine mammary epithelial cells has the beneficial effects that by the cell in-vitro culture method for primary culture of the bovine mammary epithelial cells, the purified bovine mammary epithelial cells can be obtained successfully, meanwhile, cell climbing-out time is shortened, cell proliferation speed is increased, cell growth state is good, possibility of cell pollution is greatly reduced, and the problem that beestings of mammary tissues during lactation period is not easy to clean and easy to pollute is avoided.

The invention provides a method for culturing porcine mammary epithelial cells by 3D. The method comprises the following steps: (1) preheating a pore plate at 37 DEG C; (2) digesting porcine mammary epithelial cells of generations earlier than the 10th generation into single cells by trypsin, performing centrifugation, removing supernatant, and retaining precipitate; (3) resuspending the precipitate with a mixed solution prepared from Matrigel and an Advanced DMEM / F12 complete medium at 0-10 DEG C in a volume ratio of 1:1, and adjusting the concentration of the porcine mammary epithelial cellsto 1*10<4>-5*10<4> cells / ml; (4) adding dropwise the resuspension solution according to 30-40 mul per drop to the pore plate, and inverting the pore plate for incubation at 37 DEG C for 30-40 min toform gel; (5) adding the Advanced DMEM / F12 complete medium to the pore plate for continuous culture for 2-3 days, and replacing the Advanced DMEM / F12 complete medium once every other day.

The present invention is directed to neutralizing prolactin receptor antibodies 002-H06 and antigen binding fragments, pharmaceutical compositions containing them and their use in the treatment or prevention of benign disorders and indications mediated by the prolactin receptor such as endometriosis, adenomyosis, non-hormonal female contraception, benign breast disease and mastalgia, lactation inhibition, benign prostate hyperplasia, fibroids, hyper- and normoprolactinemic hair loss, and cotreatment in combined hormone therapy to inhibit mammary epithelial cell proliferation. The antibodies of the invention block prolactin receptor-mediated signaling.

The invention discloses an immortalized dairy cow mammary epithelial cell line, a construction method, and applications thereof. In the provided construction method, Chinese Holstein cow epithelial cells are taken as the host cells, retrovirus carriers with SV-40-large T antigen are used to infect host cells, and finally a strain of immortalized dairy cow mammary epithelial cell line that can perform stable passage is screened out. The microbial preservation number of the strain is CGMCC No.8707. The constructed immortalized dairy cow mammary epithelial cell line has a stable property, the cell morphology is similar to that of primary cells after 40 generations, and the cells grow faster than the primary cells, can characteristically express mammary epithelial cell specific proteins, and maintain the synthesis functions of milk fat and milk protein. The immortalized dairy cow mammary epithelial cell line can be used as a cell model which is widely used in the fields such as milk component synthesis, milk secretion mechanism, research on laticiferous cell apoptosis and mammary gland degradation mechanism, and the like, and can be used as a mammary gland bioreactor to produce bioactive proteins.

The invention discloses a method for isolating and cultivating milk cow breast cells in vitro, relating to the cell in vitro isolation and culture technology field and solving the problem that breast fibrocytes and epithelial cells of a milk cow cannot be isolated completely during the isolation and culture of the breast epithelial cells of the milk cow. The method comprises the following steps of: selecting mammary tissue of a milk cow which is killed on the very day from a local abattoir, adopting tissue explants technique and graded trypsin digestion technique to isolate and cultivate the breast epithelial cells of the milk cow according to the different sensibility of the fibroblasts and epithelial cells to trypsin and the different adhering time, purifying the breast epithelial cells of the milk cow, identifying with keratin 18 to obtain the corresponding cell system finally. The method provides the best trypsin concentration when isolating and cultivating the breast epithelial cells of a milk cow, and can completely isolate the breast fibrocytes and the epithelial cells of the milk cow from each other.

The invention discloses a preservation method of ruminant mammary gland epithelial cells. The preservation method comprises the following steps of: (1) preparing a cryoprotective solution which is prepared from the following components in percentage by weight: 10 percent of DMSO (Dimethyl Sulfoxide), 40 percent of fetal calf serums and 50 percent of cell suspension; and (2) gradually and slowly lowering the temperature of the obtained cryoprotective solution until the temperature of the cryoprotective solution is lowered to -196 DEG C and cryopreserving the cryoprotective solution at -196 DEG C with liquid nitrogen. By adopting the method of the invention, the preservation time of the ruminant mammary gland epithelial cells can be prolonged.

The invention relates to a method for utilizing a gene chip to analyze lipopolysaccharide to stimulate variation of the expression profile of dairy cow mammary epithelial cells. The method comprises the following steps that 1, a dairy cow mammary epithelial cell inflammation model is built up; and 2, traditional Chinese medicine is tested by using the dairy cow mammary epithelial cell inflammationmodel.

The invention discloses a dairy cow mammary gland endothelial cell line under ACADSB gene knockout and a construction method of the cell line. According to the method, a recognition sequence of gRNA for knocking out the ACADSB gene is utilized, and the nucleotide sequence of the recognition sequence is shown in SEQ ID NO.1. The ACADSB gene is precisely and efficiently knocked out through the designed gRNA, and the dairy cow mammary gland endothelial cell line under ACADSB gene knockout is effectively constructed. According to the dairy cow mammary gland endothelial cell line and the construction method, the defect that an RNAi technology is low in interference efficiency is overcome, and the method for constructing the gene-knockout cell line is simple and efficient. The ACADSB gene is animportant marker gene relevant to lipid metabolism and fatty acid metabolism; through the construction of the knockout cell line of the gene, effective materials and gene tools are provided for further research on gene functions relevant to lipid metabolism and fatty acid metabolism pathways.

The invention relates to the fields of tissue and cell engineering and particularly provides a single cell cloning method for obtaining goat mammary epithetical cells. According to the method, single cell cloning preparation is performed through a micromanipulation method, tedious cell counting and repeated dilution in a traditional method are eliminated, single cell cloning operation can be accomplished within a short time, and the single cell cloning formation rate is increased. The goat mammary epithetical cells obtained through the method provided by the invention can be used for researching functions of different lineages of mammary epithelial cells and the interaction of the mammary epithelial cells during a lactation process.