65 results about "Sertoli cell" patented technology

A method of in vitro maturation of adult human germ line cells in an artificial biological environment, which entails:a) isolating human spermatogonial stem cells (SSCs), and optionally purifying the same; andb) co-culturing the isolated and optionally purified SSCs with a suitably adjusted Sertoli cell environment to obtain haploid germ cells.

The present invention discloses methods for isolating, proliferating and differentiating germ-line stem cell in vitro. More specifically, the present invention discloses a method for the in vitro isolation and proliferation of mammalian germ-line stem cells, characterized in culturing cells isolated from mammalian testis in an embryonic stem cell culture medium, and for the in vitro differentiation, characterized in encapsulating mammalian germ-line stem cells and Sertoli cells with calcium alginate and co-culturing with peritubular cells. Also, the present invention discloses germ-line stem cells obtained by above methods, a composition for the treatment of male infertility, comprising the germ-line stem cells, and a method for the treatment of male infertility by the use of the germ-line stem cells.

A cell composition composed of spermatogonial stem cells, Sertoli cells, Leydig cells and optionally peritubular cells, is provided, as is a culture composition, artificial testicular construct, hydrogel composition, and device containing the same. A method for using the device as a physiologically relevant in vitro model of human testicular function to screen compounds for pharmacological or toxicological activity is also provided.

The invention discloses an adult sertoli cell-pancreas islet compound and a preparation method for the same, which solve the problem of influence on pancreas islet transplantation effect due to the transplant dysfunction because the pancreas islet separated and purified presently generates hypoxic-ischemic and immunological rejection reactions after being transplanted and then loses the function. The preparation method comprises the following steps of: 1, separating and purifying adult pancreas islet by adopting a semi-automatic filling and continuous density gradient methods; 2, separating and purifying adult sertoli cells by adopting a two-step digestion method; and 3, constructing the pancreas islet-sertoli cell compound by adopting a specific co-culture method. The adult sertoli cell-pancreas islet compound and the preparation method for the same disclosed by the invention have the advantage that pancreas islet cells and sertoli cells are coupled into a compound, the compound is used as a transplant, and the sertoli cells are attached to the surface of the pancreas islet, so as to improve the nutritional status of the pancreas islet, inhibit rejection reaction, and increase the survival rate of the pancreas islet; the operation is simple and convenient; and the success rate is high.

The subject invention pertains to methods to enhance the therapeutic effects of cell therapy in various diseases and disorders. More particularly, the present invention provides methods of inducing systemic immune tolerance, thereby reducing or eliminating the need for systemic immunosuppression, by administering isolated Sertoli cells to an individual in need of treatment, wherein the isolated Sertoli cells are administered to the individual prior to or during administration of the therapy, e.g., cellular therapy.

This invention pertains to the discovery that stem cells (e.g., bone marrow stem cells) transplanted directly into a testicular environment are transdifferentiated into bona fide Sertoli cells, and / or Leydig cells, and / or and germ cells. This provides a mechanism for the treatment of male infertility and / or testosterone deficiency. Thus, in one embodiment, this invention provides a method of treating infertility or testosterone deficiency in a male mammal. The method typically involves implanting stem cells into the testes of the mammal whereby the stem cells differentiate into germ cells and / or Sertoli cells and / or Leydig cells thereby reducing infertility and / or testosterone deficiency.

The invention relates, in part, to non-neonatal Sertoli cells derived from non-rodent animals, pharmaceutical compositions comprising such Sertoli cells, and uses thereof. The non-neonatal, non-rodent Sertoli cells express more FasL than neonatal Sertoli cells, and they provide greater immunoprivilege than neonatal Sertoli cells. In some embodiments the Sertoli cells are modified to express a biological factor. In other embodiments, the pharmaceutical compositions further comprise non-Sertoli cells. The invention also provides implantation devices comprising the pharmaceutical compositions, methods of making the pharmaceutical compositions, and methods of using the pharmaceutical compositions by administering an effective amount of the compositions.

The present invention provides a method of providing an individual with a biological factor or intermediate thereof which comprises introducing into the individual Sertoli cells genetically altered to produce the biological factor or intermediate thereof. The genetically altered Sertoli cells are administered in an amount effective to produce the desired effect. Aside from producing the biological factor or intermediate thereof, the engineered Sertoli cells also create an immunologically privileged site. Vectors comprising a promoter which functions in Sertoli cells operably linked to coding sequence for a desired biological factor are also provided as are Sertoli cells comprising such vectors. A pharmaceutical composition comprising Sertoli cells genetically altered to produce a biological factor is also provided.

The invention relates to a method for improving the sperm motility with Sertoli cells as the trophoblast. The method mainly includes the steps of: (1) conducting isolation culture on Sertoli cells; (2) preparing a Sertoli cell trophoblast: conducting isolation culture on Sertoli cells for 2 days to form a cell monolayer, adding mitomycin C to perform treatment for 2h so as to prepare a Sertoli cell trophoblast; and (3) acquiring sperm from an epididymis of a mature male animal, adding the sperm into a culture system adopting Sertoli cells as the trophoblast, and performing co-culture on the sperm and Sertoli cells. The method provided by the invention adopts Sertoli cells as the trophoblast for in-vitro culture of sperms in testicle and epididymis, solves the problem of low in-vitro sperm motility, enhances the motility and survival rate of in-vitro sperms, and increases the fertilization capability of sperms.

The invention discloses a method for marking germ cells and Sertoli cells of spermatogenic epithelium of turbot in different developmental stages and an application. Fragment Amh / Sox9 / Gsdf is amplified from cDNA of turbot spermary, cloning, preferring and plasmid extraction are performed sequentially, plasmids are linearized by restriction enzymes, probes are prepared after plasmid recovery, fixation, dehydration and entrapment are performed after sample processing, fluorescence in situ hybridization is performed after slicing, finally, nuclear staining is performed, and slice sealing, observation and photographing are performed. The method has the beneficial effects as follows: spermatids and the Sertoli cells of the spermatogenic epithelium of the turbot in different developmental stagesare separated out skillfully, and the marking method is simple and feasible and higher in precision; the method for marking the germ cells and the Sertoli cells of the spermatogenic epithelium of theturbot in different developmental stages can be applied to distinguishing and marking of cells of the spermatogenic epithelium of other marine organisms, and an effective method and a scientific basis are provided for studying spermatogenesis of fish and environment in the spermatogenic epithelium in future.

The invention discloses a natural passage cell line for lamb testis sertoli cells and application of the natural passage cell line to separate culture and proliferation of goatpox viruses. The cell line disclosed by the invention is a lamb testis sertoli natural passage cell line which is obtained by separating and purifying primary cells of a lamb testis to obtain the lamb testis sertoli cells and performing natural passage on the lamb testis sertoli cells. A research shows that the cell line is relatively sensitive to inoculation of the goatpox viruses; the virus titer proliferated by the cell line is greater than that of the primary cells, Vero cells and BHK21 cells of the testis of a newborn lamb by about 1.5, 2.5 and 2.3 respectively. The cell line for separating the goatpox viruses can guarantee the uniformity and the stability of the viruses and prevent the risk that the separated viruses carry other pathogenies due to the fact that the primary cells are used for multiple times. Furthermore, by the use of the passage cell line for preparing vaccines, the production process is simplified, the production period is shortened, and the production cost is reduced; meanwhile, the quality of the vaccines are kept stable; the cell line has certain practical significance for large-scale production of the vaccines.

According to the present invention, there is provided a biological chamber system having a biochamber defined by outer walls of Sertoli cells. Also provided is a transplantation facilitator including a biochamber. A method of making biochambers by co-culturing facilitator cells and therapeutic cells and then aggregating the facilitator cells is also provided. Also provided is a method of transplanting cells by incorporating transplant cells into a biochamber and transplanting the biochamber containing the transplant cells.

The invention discloses a biological sponge based on acellular dermal matrix and a preparation method thereof. The biological sponge is made of an acellular dermal matrix as a raw material, and Ca2<+>and fa-PEG are used for cross-linking. The biological sponge prepared by the invention has a porous structure, has pores which communicate with one another, and has a three-dimensional structure similar to human skin tissue, can enable body tissue cells to grow rapidly, has good plasticity, and can be prepared into different shapes as needed so as to meet the needs. The biological sponge of the invention is an advanced tissue engineering scaffold material, contains rich growth factors, has good biocompatibility, is more conducive to the growth of Sertoli cells, can not only be used for woundrepair, but also is an excellent tissue engineering skin scaffold material, and can be used for repairing skin defects and preparing tissue engineering skin.

The invention discloses a bovine testis sertoli cell carcinoma cell, and application thereof in separation and culture of poxviruses. The bovine testis sertoli cell carcinoma cell is named as a bovinetestis sertoli cell carcinoma cell BTSCC, and the preservation number of the bovine testis sertoli cell carcinoma cell BTSCC is CCTCC NO:C201931. Furthermore, the invention also provides applicationof the bovine testis sertoli cell carcinoma cell in separation and culture of poxviruses such as ORFV and GTPV. The cell is sensitive to ORFV and GTPV cytotoxicity, can efficiently proliferate the ORFV and GTPV viruses, and can ensure the uniformity and stability of the viruses. The titer of the virus proliferated by the cell is equivalent to that of the newborn bovine testicular sertoli cell (NBSC), and the bovine testis sertoli cell carcinoma cell is high in growth speed and very high in seed division rate and can grow under a low-serum culture condition. The ORFV and GTPV viruses are separated and cultured by using the cell, so that the test efficiency can be improved, the test cost is reduced, and meanwhile, the quality stability of vaccines prepared from the cell is ensured. A new technical means is provided for poxvirus culture and large-scale production of poxvirus vaccines.

The invention discloses an anti-ANGPTL3 (angiopoietin-like protein) monoclonal antibody. The heavy chain amino acid sequence of the monoclonal antibody is SEQ ID: No 1, the light chain amino acid sequence of the monoclonal antibody is SEQ ID: No 2, the heavy chain DNA (deoxyribonucleic acid) sequence of the monoclonal antibody is SEQ ID: No 3, and the light chain DNA sequence of the monoclonal antibody is SEQ ID: No 4. The invention also discloses application of the anti-ANGPTL3 monoclonal antibody in preparing a medicine capable of treating a nephrotic syndrome. The anti-ANGPTL3 monoclonal antibody specifically identifies an ANGPTL3, blocks a fibrinogen-like structural domain of the ANGPTL3, and antagonizes the injury of the ANGPTL3 to sertoli cells. The invention can be used for detecting the ANGPTL3 and is hopefully used for treating nephrotic syndrome proteinuria.

Fertility management can include: administering to the subject one or more doses of a compound according to Formula I so as to reduce fertility in the subject. Fertility management can also include administering an effective amount of the compound to: impair Sertoli cell function in a male subject; inhibit spermatogenesis in the subject; reduce testis weight in the subject; reduce ovary weight in a female subject; reduce serum progesterone in the female subject; impair ovarian follicle function in the female subject; causing reversible fertility in the subject. In order to return fertility, the method can include ceasing administration of the compound to the subject so as to return fertility in the subject. The compound can be administered for irreversibly sterilizing the subject.

The present invention provides a method of producing a non-human vertebrate that harbours germ cells having a desired gene transferred thereto, comprising injecting the desired gene to the testis of a non-human vertebrate wherein no tight junction exists between Sertoli cells to transfer the desired gene to germ cells, so as to obtain a non-human vertebrate that harbours the germ cells having the desired gene transferred thereto. Using the method of the present invention, even in animal species and lines for which in vitro transduction has been difficult to date, it is possible to obtain an individual harbouring germ cells, particularly spermatogonial stem cells, having a desired gene transferred thereto, at extremely high efficiency. Also, the fertility of the male to receive an injection of the gene is retained, compared to in vitro transduction of germ cells, because gene transfer is achieved without reducing the number of spermatogonial stem cells in the testis and transgenic sperms and transgenic animals can easily be prepared.

The invention discloses a method to reprogram fibroblasts into Sertoli cells in vitro and application of the method. The method comprises the steps of introducing into fibroblasts, a fluorescent protein reporter system that specifically expresses in Sertoli cells; introducing a substance that increases NR5A1 protein expression and / or activity; adding G418, and culturing for 3-7 days; discarding anaqueous phase, adding a medium to culture fibroblasts, and continuing to culture for 3-7 days; isolating Sertoli cells from the culture system. Experiments prove that the cells prepared by the abovemethod fully attain the characteristics of Sertoli cells, such as ability to attract endothelial cells, ability to form lipid droplets, ability to interact with reproductive cells, ability to inhibitthe growth of lymphocytes and occurrence of interleukin, and ability of immune privilege and the like. The method has important applicable value in the fields, such as treatment of infertility, allograft, and cell therapy.

Aggregates and their method of preparation suitable for implantation into a recipient in order to produce insulin in vivo. The methods involve culturing islet cells isolated from the pancreas of donor piglets with isolated Sertol i cells from the testes of donor piglets. A preferred period of culturing is 5 days and may be followed by a purification procedure.

The invention discloses a detection method of a protein marker for urine exosome. The method comprises the following steps: extracting and purifying exosome from freshly collected urine, collecting precipitate, and performing resuspending, so as to obtain exosome suspension; performing separation on the exosome suspension obtained in the step (1) through magnetic beads coated by CD13 and podocalyxin respectively, so as to obtain an exosome separation fluid; measuring the protein concentration of exosome of the separation fluid through a BCA kit; preparing the separation fluid into a protein loading sample according to the protein concentration, and performing CD13, Podocalyxcin and CD63westernblot detection on the sample, so as to obtain the required protein marker after identification. The method has the advantages that positive exosome of podocalyxin and CD13 in urine can be respectively extracted, and the method is beneficial to further analysis and detection to the exosome, so thatchange of sertoli cell and proximal renal tubular epithelial cell can be accurately reflected, and a biological marker is provided for diagnosis and treatment of diseases.

The present invention provides a method of producing a non-human vertebrate that harbours germ cells having a desired gene transferred thereto, comprising injecting the desired gene to the testis of a non-human vertebrate wherein no tight junction exists between Sertoli cells to transfer the desired gene to germ cells, so as to obtain a non-human vertebrate that harbours the germ cells having the desired gene transferred thereto. Using the method of the present invention, even in animal species and lines for which in vitro transduction has been difficult to date, it is possible to obtain an individual harbouring germ cells, particularly spermatogonial stem cells, having a desired gene transferred thereto, at extremely high efficiency. Also, the fertility of the male to receive an injection of the gene is retained, compared to in vitro transduction of germ cells, because gene transfer is achieved without reducing the number of spermatogonial stem cells in the testis and transgenic sperms and transgenic animals can easily be prepared.

The invention relates the method for checking laver raw sugar to hold cell proliferation action for mouse testis. Now there are no related reports. The invention uses MTT method to check the breeding rate, and the results express that the more the raw sugar, the stronger the breeding action. The invention can hold cell to graft together with other cell and improve the spermium quality.

The invention discloses a newborn sertoli-cell immortalization cell line and an establishing method and application thereof. The newborn sertoli-cell immortalization cell line is named as the newbornsertoli-cell immortalization cell line hTERT-NBSC, and the collection number of the hTERT-NBSC is CCTCC NO:C2018201. Tests find that the hTERT-NBSC can continuously pass 65 generations or above, and the growth and proliferation characteristic is kept constant. The cell line is sensitive to inoculation of ORFV, the ORFV virus can be efficiently proliferated, and the uniformity and the stability ofthe virus can be guaranteed. The titer of the ORFV virus proliferated by the cell line is equivalent to that of the newborn sertoli cell, the cell sorting rate of the ORFV virus is higher than that ofthe NBSC, and the cell sorting rate can be 1:3. Meanwhile, vaccine is prepared with the passage cell line, the production technology is simplified, the production cycle is shortened, the production cost is reduced, and the stable quality of the contagious ovine ecthyma vaccine prepared with the method is also guaranteed. Therefore, through the newborn sertoli-cell immortalization cell line and the establishing method and application thereof, a new technological mean is provided for large-scale production of the contagious ovine ecthyma vaccine.

Fertility management can include: administering to the subject one or more doses of a compound according to Formula I so as to reduce fertility in the subject. Fertility management can also include administering an effective amount of the compound to: impair Sertoli cell function in a male subject; inhibit spermatogenesis in the subject; reduce testis weight in the subject; reduce ovary weight in a female subject; reduce serum progesterone in the female subject; impair ovarian follicle function in the female subject; causing reversible fertility in the subject. In order to return fertility, the method can include ceasing administration of the compound to the subject so as to return fertility in the subject. The compound can be administered for irreversibly sterilizing the subject.

The invention relates to the technical field of a traditional Chinese medicine preparation, and concretely relates to a traditional Chinese medicine composition for preventing and treating diabetic nephropathy and a preparation method thereof. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 550-650 parts of radix astragali, 150-250 parts of root of kudzu vine, 150-250 parts of eucommia, 150-250 parts of gotu kola, 100-200 parts of Periostracum Cicadae, 100-200 parts of white batryticated silkworm, 100-200 parts of arctium fruit, 40-80 parts of rheum officinale, and 100-200 parts of turmeric. The composition is precise and appropriate, the curative effect is obvious, the raw materials can be combined for usage, acrid opening and bitterdownbearing are realized, the composition can invigorate and enhance the human body quality at the same time, and the functions of tonifying kidney and dispersing blood stasis, and enlightening mysterious mansion can be achieved. The traditional Chinese medicine composition has good curative effect for reducing urine protein and improving kidney function, can improve DN general symptom, protects kidney function, delays DN progress, and is related to expression of up-regulated sertoli cell associated protein nephrin and podocin.

Technology for the isolation and propagation of primary human Sertoli cells from normal testes tissue, including cultures of proliferative primary human Sertoli cells for research and clinical applications, and a pharmaceutical composition for cell therapy, ex vivo gene therapy, and for the reduction of autoimmune, allograft, and xenograft immune reactions.

A cell composition composed of spermatogonial stem cells, Sertoli cells, Leydig cells and optionally peritubular cells, is provided, as is a culture composition, artificial testicular construct, hydrogel composition, and device containing the same. A method for using the device as a physiologically relevant in vitro model of human testicular function to screen compounds for pharmacological or toxicological activity is also provided.

The invention discloses application of resveratrol to treating kidney disease. Animal experiment research shows that the resveratrol has better function of protecting the kidney, remarkable inhibition function of pathological symptoms of proteinuria, sertoli cell injury, mesangial cell proliferation, extracellular matrix accumulation and the like, and better prevention and treatment functions on primary nephrotic syndrome and mesangial proliferative glomerulonephritis. The resveratrol has easy obtaining of raw materials, strong pathological activity and low adverse effects, and is hopeful to be a new generation medicine for treating the kidney disease.

The invention discloses a method for inducing pluripotent stem cells by bovine sertoli cells, and belongs to the field of stem cells. The method comprises the following steps: step 1, separating bovine sertoli cells; step 2, cultivating the bovine sertoli cells; and step 3, transferring four transcription factors encoding Oct4, Sox2, c-Myc and Klf4 into the bovine sertoli cells so as to produce the pluripotent stem cells. Compared with the prior art, the method fully utilizes the characteristics of differential adherence of the bovine sertoli cells, and cultivates the bovine sertoli cells withuniform morphology. Since the bovine sertoli cells are derived from meat calf, the bovine sertoli cells have a high reprogramming ability. The bovine sertoli cells replace bovine adult fibroblasts and mesenchymal stem cells, and are used as source cells for preparing and inducing pluripotent stem cells, so that not only the cell acquisition is simple, but also the reprogramming rate is high.