38 results about "Leydig cell" patented technology

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 provides an external directional induced differentiation method for differentiating human induced pluripotent stem (iPS) cells into leydig cells (LCs) through neural crest stem cells (NCSCs). By the utilization of an animal model, it is verified that the LCs from the iPS cells have the capacity of regenerating aged or damaged LCs, and a new testosterone-supplying scheme is provided for a patient with hypogonadism and particularly an LOH patient.

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 discloses a method for obtaining Leydig cells and purposes of the method. The method disclosed by the invention has the advantages of quickness, simpleness and convenience, and capability of obtaining a large quantity of Leydig cells with high purity and good functions, and is used for establishing seed cells of tissue engineered androgen-secreting tissue and other purposes.

The invention discloses a method for inducing human ADSC (Adipose-Derived Stem Cells) to be differentiated into LCs (Leydig Cells) by micromolecules. The method comprises the following steps: firstly,separating and extracting the human ADSC, and completing identification; then, inducing differentiation of the human ADSC by adopting a specific culture medium, and simultaneously adding a certain amount of micromolecules such as SAG, 22OHC, Li, PDGF-AA, FGF2, Androgen and LH in the special culture medium at different time points, thus promoting differentiation; finally, manually removing ALCs (Adult Leydig Cells), enriching and differentiating target cells ADSC-ALCs, and completing the identification. By inducing according to the method, the human ADSC can be successfully differentiated intothe LCs, so that a cell source is provided for treating testosterone deficiency by adopting cell transplantation.

The present invention encompasses methods and compositions for reducing an immune response to a transplant in a recipient by treating said recipient with an amount of liver stromal cells effective to reduce or inhibit host rejection of the transplant. Also disclosed is a method of inducing a reduced immune response against a host by foreign tissue, i.e., graft versus host disease, by treatment with liver stromal cells.

The invention discloses a method for constructing a machin progenitor leydig cell immortal line. The method comprises the following steps: by taking machin testicular tissues, obtaining leydig cells of a high ratio, culturing in a complete medium for 20-28 hours, replacing fresh culture solution, continuously culturing, and performing cell passage to the leydig cells presenting fibroblast growth in uniform form; infecting the leydig cells by utilizing slow virus solution carrying Large T and Egfp, performing fluorescence labeling cell screening, and performing cell passage to a cell strain for stably expressing Egfp; and screening the cell strain for stably expressing Egfp, identifying the progenitor leydig cells, continuously performing cell passage for over 50 generations, and finally constructing the machin progenitor leydig cell immortal line. The in-vitro cell passage of the progenitor leydig cell immortal line with the transferred Large T exceeds 62 generations, the leydig cell labeled proteins such as CYP11A1, CYP17A1 and STAR are expressed, marker genes of mesenchymal stem cells such as Pdgfral and Lifr are expressed, the cell shape is maintained into the fiber shape, and a characteristic progenitor leydig cell immortal line is obtained.

The invention discloses a method for testing that SLC can maintain homeostasis of Leydig and muscle-like cells in adult testis. Stem cells related to testicular seminiferous tubules of adult SD rats are called as Leydig stem cells (SLC) and can be differentiated into Leydig cells in vitro and in vivo. Proliferation and differentiation of the SLC are regulated by locally generated factors, including PDGF family members PDGF-AA and PDGF-BB. Both members are able to stimulate proliferation of SLC, while only PDGF-AA is able to stimulate differentiation of SLC into Leydig cells producing testosterone (T), while PDGF-BB inhibits the process. In the method, it is verified that the SLC can actually form Leydig cells or myocyte-like cells under different differentiation conditions. PDGF-AA + LH can enable the SLC to be differentiated into Leydig cells capable of generating T, and PDGF-BB + TGFB induces the cells to become muscle-like cells for expressing smooth muscle actin. The method can effectively verify pluripotency of seminiferous tubule-associated stem cells (SLC), and test important role of SLC in maintaining homeostasis of Leydig and Myoid populations in vivo.

The invention relates to a method for inducing adipose tissue-derived stem cells to differentiate into testicular interstitial-like cells by a single transcription factor, and belongs to the fields of stem cell biology and regenerative medicine. According to the method, chemically modified mRNA of a transcription factor NR5A1 is synthesized in vitro, exosomes extracted from autologous urine of a patient are used as a delivery carrier to form an exosome-modNR5A1 compound, and then the exosome-modNR5A1 compound and ADMSCs derived from autologous tissue of the patient are co-cultured to induce the ADMSCs to be differentiated into Leydig-like cells with a testosterone secretion function. According to the method disclosed by the invention, the Leydig-like cells with the testosterone secretion function can be quickly and efficiently obtained, meanwhile, the cells can effectively react to the stimulation of luteinizing hormone, and the cells have a gene expression profile related to the androgen synthesis of mature Leydig cells. The Leydig cell obtained by the invention has a good application prospect in the aspect of treatment of male hypogonadism.

The use of an aromatase blocker or an estrogen blocker is described in a method for increasing spermatogenesis and Seritolli cell function, and / or improving Leydig cell function, in order to to increase endogenous testosterone levels in a male mammal. The levels of active materials used are significantly lower than the levels of these materials used to treat female estrogen sensitive tumors.

The partial androgen deficiency of the aging male (PADAM) and the supplement of synthetic testosterone is an important research subject in the American biological science. The activity of leydigs cells of testis and the synthesizing and excreting state of testosterone has aroused wide concern of the medical circles. Taking synthetic testosterone may cause the cancer of genital system, which makes the middle-aged and old male people do not dare to taking the drugs of the synthetic testosterone. The Compound Recipe Product of 4 Kinds of Plants to Increase Testosterone is made by following procedures: Under the carefully and strictly controlled condition, use the technology of absorption and elution of WLD resin to extract and purify it and measured by the gas chromatograph and make a fingerprinting chromatograph quantitative determination so as to guarantee the product has a stable technological process and a high quality. The experiment of pharmacological activity shows that, by activating the leydigs cells of testis to enhance its ability to excrete testosterone, the compound recipe product has a remarkable function to obviously raise the quantity of testosterone blood serum of the normal and injured genital system and is used for treating the partial androgen deficiency of the aging male (PADAM) in clinical treatment.

The invention discloses a method for culturing high-purity primary Leydig cells of Guizhou fragrance pigs. The method is characterized by comprising the following steps: (1) digesting testicular tissue blocks of fragrance pigs through collagenase to obtain a cell suspension solution; (2) repeatedly digesting with 0.25 percent Trypsin-EDTA to obtain Leydig cells; (3) enriching the Leydig cells through a differential centrifugation method, wherein the differential centrifugation method at least comprises three times of centrifugation, the speed of the first-time centrifugation is 800 to 1200 rpmand the time is 3 to 8 min, and the speed of the second-time and third-time centrifugation is 600 to 1000 rpm and the time is 3 to 8 min; (4) purifying the Leydig cells through a differential attachment method. Compared with an existing primary culture and purification method of pig Leydig cells, the method has the advantages that the operation is simpler and more efficient, and the Leydig cellsof the Guizhou fragrance pigs, which have a great quantity, strong activity and high purity, are obtained. The method disclosed by the invention can provide effective technological guidance for deeplyresearching sexual precocity properties of the Guizhou fragrance pigs and related functions of the Leydig cells.

The invention discloses a long-term culture method for swine testicle mesenchymal progenitor cells. The long-term culture method includes separation, detection and differentiation in vitro, and specifically includes the steps of culturing piglet testicle fibroblasts for 5-8 days by means of an in-vitro proliferation culture medium of swine PLCs (progenitor leydig cells) to acquire a PLCs clone colony, culturing by means of a differentiation medium of swine PLCs so that PLCs begin to differentiate into mature testicle mesenchymal cells in the third day, then culturing in the proliferation culture medium, and maintaining in vitro for six month at most. According to the long-term culture method, a mature process of separation, culture, detection and differentiation of swine PLCs is created initially, and proliferation and differentiation maintained in vitro of PLCs can be achieved and controlled by different in-vitro culture systems determined finally. The long-term culture method has high application value in research on biological property of PLCs in vitro, research and development of drugs and vaccine production and the like.

The invention discloses a biomarker of perimuscular cells of testicular tubes and application of the biomarker. The biomarker comprises an ITGA9 protein. On the basis of the ITGA9 protein, the kit for identifying the testicular perimuscular cells is prepared, the anti-ITGA9 protein antibody is used for specifically recognizing and combining the perimuscular cells, then the testicular perimuscular cells can be effectively identified through methods such as flow analysis and the like, in addition, after the testicular perimuscular cells are identified, the testicular perimuscular cells can be distinguished from Leydig cells, and therefore the identification accuracy of the testicular perimuscular cells is improved. Therefore, the two cells can be effectively separated, the problem that the two cells are difficult to separate by conventional stepwise digestion, differential adherence, gradient centrifugation and other methods is solved, and high-purity Leydig cells and high-purity peritubular muscle-like cells can be obtained through separation and purification.

The invention discloses a method for fully utilizing small molecules to induce human fat stem cells to differentiate into Leydig cells (LCs). The steps of the method are as follows. First, human adipose stem cells are isolated and identified; then, a specific medium is used to induce the differentiation of human adipose stem cells, and a certain amount of small molecules are added to the medium at different time points, including: SAG, 22OHC, Li, PDGF‑ AA, FGF2, Androgen, and LH promote differentiation; finally, the non-testis Leydig cells (ALCs)-like cells are manually removed to enrich the differentiation target cells (ADSC‑ALCs), and the identification is completed. Induced according to the method of the present invention, human adipose stem cells can be successfully differentiated into Leydig cells of the testis, thereby providing a cell source for the treatment of testosterone deficiency by cell transplantation.

The invention relates to medical application of ethane dimethane sulfonate (EDS). EDS can be used for eliminating leydig cells and preparing a medicine for treating and / or preventing leydig cell dependent diseases. When low-dosage EDS is injected into testicular arteries, the concentration of testosterone can achieve and be kept below a castrating level; furthermore, systemic side reaction nearly does not exist; health is not influenced; toxicity is also not generated; EDS is sensitive on human leydig cells and has high safety.

The invention discloses a method for separating rat testis interstitial Leydig cells. The method comprises the following steps: acquiring a rat testis and preparing a rat testis cell suspension; marking rat testicular cells; screening positive cells by adopting a magnetic bead sorting (MACS) method; and repeatedly purifying the positive cells through adoption of a magnetic bead sorting method. The separation method disclosed by the invention is simple and easy to operate, high in cell yield, high in purity and high in activity, and does not need expensive equipment.

The invention relates to a gene medicine for treating testicular interstitial cell dysfunction and an application thereof. The gene medicine is a recombinant adeno-associated virus capable of expressing a luteinizing hormone / chorionic gonad hormone receptor by using host cells. The medicine not only can recover the testosterone level and promote gonad development, but also can obviously promote sperm generation and rebuild fertility; and meanwhile, the gene medicine has no obvious side effect and is high in safety. In addition, the gene medicine can be applied to preparation of gene medicine compositions or bioreactors for producing luteinizing hormone / chorionic gonad hormone receptors, and has very high application value.

The present application provides an in-vitro committed differentiation method for inducing human induced pluripotent stem cells (hiPSCs) into Leydig cells (LCs) by neural crest stem cells (NCSCs). The hiPS-derived LCs is verified by an animal model to have the capacity of regenerating senile or injured LCs, so that a new treatment for supplementing testosterone is provided for patients suffering from hypogonadism, particularly for patients suffering from late-onset hypogonadism (LOH).

The invention relates to an isolated culture method and application of tree shrew testicular interstitial cells. The method comprises the steps of separation and purification. According to the invention, the isolated culture method of the tree shrew testicular interstitial cells is successfully established, the susceptibility and virus proliferation characteristics of Zika virus to the tree shrew testicular interstitial cells are determined, and a basis is provided for research on a Zikv invasion reproductive system mechanism.