106 results about "Sertoli cells" patented technology

Methods for therapeutically programming human adult stem cells into pluripotent cells are provided. Cell therapeutically programmed from adult testicular cells are disclosed. The therapeutically reprogrammed cells are suitable for cellular regenerative therapy and have the potential to differentiate into more committed cell lineages.

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 invention relates to a porcine transmissible gastroenteritis and epidemic diarrhea combined live vaccine and a preparation method of the porcine transmissible gastroenteritis and epidemic diarrhea combined live vaccine. The porcine transmissible gastroenteritis and epidemic diarrhea combined live vaccine is prepared by performing virus amplification on a swine testicular cell line (ST cells) or an African green monkey kidney cell line (Vero cells) by using a self-attenuated and preserved transmissible gastroenteritis virus SD / L strain and a self-attenuated and preserved porcine epidemic diarrhea virus LW / L strain, and carrying out the steps of harvesting, uniformly mixing, freeze-drying and the like. The porcine transmissible gastroenteritis and epidemic diarrhea combined live vaccine can effectively prevent two diseases namely swine transmissible gastroenteritis and epidemic diarrhea.

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 yolk antibody for pig's infective enterogastritis virus is prepared through reproducing the said virus on pig's testis cell, applying it as immunogen to the layer, collecting yolk and purifying. When it is applied to pig, the comparison between experimental and reference groups shows that it has high effect on preventing and cuting pig's infective enterogastritis.

The invention relates to an ST cell adapting to full-suspension culture, and an application in vaccine virus culturing, and belongs to biotechnical field. The ST cell adapting to full-suspension culture is named as a swine testicle cell suspension adaptation strain ST-2014S, and is preserved in China Center for Type Culture Collection on May 13, 2015 with the preservation number of CCTCC C201567. The invention also discloses a method for culturing a vaccine virus through using the full-suspension adaptation ST cell. Compared with vaccine viruses in the prior art, the full-suspension adaptation ST cell cultured vaccine virus has the advantages of high automation degree, no need of vector intervention, realization of suspension culture in a serum-free or low-serum medium, meeting of large-scale industrial requirements of virus culture, development of a large-scale production method meeting GMP production technology requirements, and very good industrial application prospect.

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 methods for culturing a swine testis cell in a serum-free manner and producing a classical swine fever cell vaccine by using the swine testis cell. The method for culturing the swine testis cell in the serum-free manner comprises the following steps: culturing the swine testis cell by a culture medium containing 10% calf serum; when the cell grows to 60%, changing a culture medium containing 5% calf serum to carry out adaptive culture; when the cell is dead and is suspended, replacing a new culture medium; when the cell completely grows, carrying out subculture by using a culture medium containing 10% calf serum for 3-5 generations; repeating the steps, and gradually reducing the serum content to 5%, 2% and 1% from 10% till realizing serum-free culture; and finally, screening a pure cell line by using a disappearing dilution method. According to the methods, the swine testis cell can be obtained by serum-free habituated culture; the swine testis cell can enable classical swine fever viruses to be multiplied and is applied to the production of a classical swine cell vaccine, thus not only is the risk caused by use of animal serum avoided, but also the components of the culture mediums are clear, the identity among batches is improved and the downstream purifying process is simplified.

Cervical cancer cells and HPV+ head and neck cancer cells express three testis-specific genes not normally expressed in somatic cells: testicular cell adhesion molecule 1 (TCAM1), synaptonemal complex protein 2 (SYCP2) and stromal antigen 3 (STAG3). Among the three markers, TCAM1 and SYCP2 are early detection markers. Various methods for identifying a human or non-human animal as a candidate for further examination for cervical cancer, preneoplastic lesion for cervical cancer, head and neck cancer, or preneoplastic lesion for head and neck cancer are disclosed. Methods of detecting said cancers and preneoplastic lesions, methods of screening for drugs for treating said cancers and preneoplastic lesions, methods for monitoring the effectiveness of a treatment for said cancers, and methods of treating said cancers are also disclosed. Further disclosed are kits that can be used to practice the above methods.

The invention relates to an ORFV virulence weakening method. A calf testicle cell line is utilized to carry out continuous passage culture of the ORFV, so that the ORFV virulence is weakened, thereby providing a stable and reliable method for preparing and producing an ORFV virulence weakening vaccine; and the method is used for obtaining abundant stable vaccines. The method disclosed by the invention comprises the following steps: 1) separating and initially culturing the ORFV; and 2) carrying out continuous passage culture on the ORFV in the calf testicle cell line until the virulence is weakened. In the step 1), the initial culture of the ORFV adopts the calf testicle cell line, and the culture fluid of the calf testicle cell line is M199, DMEM (Dulbecco Modified Eagle Medium) (high-sugar) and RPMI-1640 culture media. In the step 1), the ORFV is separated from lip scab of a sicken goat. In the step 2), the calf testicle cell line is prepared by introducing a telomerase reverse transcriptase (hTERT).

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 invention discloses a separation and culture method and application of human testis mesenchymal stem cells. The separation method comprises the steps that an obstructive azoospermia patient testis specimen which is processed through tissue biopsy separation is digested with type IV collagenase, digested cell suspension is filtered to remove tissue clumps, and human testis cells are obtained; the human testis cells are marked with a CD51 antibody, separation is performed with a flow cytometer, the cells expressing the CD51 are collected, and then the human testis mesenchymal stem cells with the positive CD51 are separated. The invention further provides the human testis mesenchymal stem cells which are obtained through the separation method and have the positive CD51, the culture method of the stem cells and application of the stem cells in preparation of medicine for treating related diseases caused by low testosterone level.

Compositions and methods are provided for eliciting antigen-specific T-cell responses against human cyclin A1 (CCNA1), which is herein identified as a leukemia-associated antigen based on its overexpression in acute myeloid leukemia (AML) including leukemia stem cells (LSC) and in immunologically privileged testis cells, but not in other normal cell types. CCNA1-derived peptide epitopes that are immunogenic for T-cells including CTL are disclosed, as are immunotherapeutic approaches using such peptides for vaccines and generation of adoptive transfer therapeutic cells.

The invention provides a bovine testicle cell line and an establishment method thereof. The establishment method comprises the following main steps: (1) primary culture, namely collecting a bovine testicle of a newborn calf in a sterile manner, cleaning adhesive tissues, separating by virtue of a trypsinization method so as to obtain bovine testicle cells with high vitality, culturing the separated bovine testicle cells with high vitality in a DMEM (Dulbecco modified eagle medium), and culturing at 37 DEG C under the condition of 5% CO2 to obtain primary bovine testicle cells; (2) transfection and screening, namely extracting and purifying eukaryotic expression plasmids pCI-neo-hTERT containing hTERT genes, leading the extracted and purified eukaryotic expression plasmids pCI-neo-hTERT to the primarily cultured bovine testicle cells in the step (1) to perform transfection by adopting a lipidosome transfection method; (3) screening and enlarged culture. The bovine testicle cells provided by the invention are easy in culture and relatively fast in growth speed, can be stably inherited, are high in motility rate and purity, and can be well preserved; the method provided by the invention is simple and convenient in operation and convenient in popularization and application. The cell line can be used for producing hog cholera vaccines, sheep poxes, ecthyma vaccines and the like.

The invention discloses a method for efficiently separating mouse spermatogonial stem cells. The method includes the following steps that firstly, mouse testis are collected, and a testicular cell suspension is prepared through a two-step enzyme digestion method; secondly, the testicular cell suspension is inoculated in a porous plate according to a certain density, and a spermatogonial stem cell culture solution is added for in-vitro culture; thirdly, the culture solution is removed, testis cells of different times are cultured through tryptic digestion, and spermatogonial stem cells are sorted from the testis cells through the magnetic activated cell sorting; fourthly, the sorted spermatogonial stem cells are inoculated to an STO trophoblast, and a spermatogonial stem cell culture solution is added for primary culture and subculture of the spermatogonial stem cells. A mouse spermatogonial stem cell system is built through the method including the steps of separation, culture, re-separation and re-culture. The method can achieve the aim of efficiently enriching the spermatogonial stem cells in vitro, and is economical and easy to implement, so that a direction is provided for protection of rare animal varieties and treatment of male infertility.

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 provides a highly classical swine fever virus infective swine testicular clone cell strain ST-B2, which has a preservation number of CCTCC NO.C2011101, and a preparation method of the swine testicular clone cell. The method comprises the steps of: 1) subjecting a swine testicular cell to limited dilution, conducting cell cloning and subcell cloning, thus obtaining a subcellular clone strain; and 2) selecting a subclone strain of the swine testicular cell with a highest classical swine fever virus infection rate, i.e. the highly infective swine testicular clone cell of the classical swine fever virus. The invention also provides a method for preparation of a high titer classical swine fever virus solution and a classical swine fever live vaccine from the swine testicular clone cell. The swine testicular clone cell provided in the invention has a high classical swine fever virus infection rate, and the classical swine fever vaccine produced from the swine testicular cell line ST clone cell strain ST-B2 by a virus-carrying and virus transmission technique has high virus titer, hard exposure to BVDV (bovine viral diarrhea virus) pollution and good pureness. By the virus-carrying and virus transmission technique, a resurgent cell can undergo continuous passage to at least 15 generations. The cell resurrection and virus inoculation times can be reduced, the production process is simplified, and the production efficiency is improved.

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.