56 results about "Testicular Interstitial Cells" 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 a method for constructing a model of oriented differentiation of embryonic stem (ES) cells into testicular interstitial cells. The method comprises the following steps of: firstly, cotransfecting a steroidogenic factor 1 and a luteinizing hormone receptor plasmid into an ES cell; then inducing the ES cell by a compound or growth factor to directionally differentiate the ES cell into the testicular interstitial cell. In the invention, the construction and the application of the model for in-vitro oriented differentiation of the stem cells into the testicular interstitialcells are demonstrated at the cellular level; a great deal of biology information is discussed initially; the constructed cell model is a substituted model for researching the influencing factor of the secretory regulation function of the testicular interstitial cell and can be used for initially selecting and evaluating the medical effect of the pharmaceutical drug using the testicular interstitial cell as the target cells; the transplanting of the testicular interstitial cell differentiated from the stem cells is used as a new method for supplying testosterone, and the application prospect of the renewable alternative treatment of the testicular interstitial cell is improved.

The invention discloses a method for induced differentiation of testicular interstitial cells and applications of the testicular interstitial cells in sexual function recovery, belonging to the technical field of biological medicines. Mesenchymal stem cells are utilized for induced differentiation of testicular interstitial cells, the testicular interstitial cells and other ingredients with a certain ratio form a medicine for sexual function recovery, the medicine for sexual function recovery is prepared into an injection, the injection is injected to the focus area in the injection form, thetestosterone level in the body is recovered in a manner that the testicular interstitial cells secrete testosterone, and thus the sexual function of the male is effectively recovered. In short, the method is safe and effective, and is strong in practicability, testicular interstitial cells are efficiently obtained through induced differentiation, and the medicine can effectively recover the sexualfunction of the male.

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 differentiating inducible pluripotent stem cells into testicular interstitial cells by small molecule induction. The method comprises the following steps: firstly,obtaining human inducible pluripotent stem cells; secondly, carrying out pretreatment culture on the human inducible pluripotent stem cells by using an E7 culture medium without basic fibroblast growth factors; thirdly, inducing the human inducible pluripotent stem cells to differentiate by using a differential medium and combining with a small molecule inducer, wherein the small molecule inducercomprises a DHH agonist SAG, 22R-OHC, lithium chloride, human platelet-derived growth factor AA, fibroblast growth factor 2, insulin-like growth factor 1, androgen, luteinizing hormone, retinol and 8-bromo-cyclic adenosine monophosphate; fourthly, manually removing clonal cluster-like parenchyma cells; fifthly, continuously culturing the remaining cells after the parenchyma cells are removed by using an enriched medium, replacing the enrichment medium regularly, and finally obtaining the target testicular interstitial cells.

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 invention discloses a method for detecting trace luteinizing hormone. The method comprises the following steps: (1) preparation of luteinizing hormone free serum; (2) preparation of testicular interstitial cell; (3) analysis on luteinizing hormone in a serum sample of a detected person; and (4) back calculation of a detection result of testosterone on a luteinizing hormone standard curve, andcheck on concentration of luteinizing hormone in the sample to be detected. The method can detect LH with extremely low concentration, has greatly improved sensitivity, is 100 times more sensitive than the existing testicular cell radio-immunity method, and is 1,000 times more sensitive than the chemiluminesence immunoassay.

The invention relates to a method for inducing mesenchymal stem cells to be directionally differentiated into Leydig cells and application of the method. Further, the invention relates to a composition for in-vitro inducing mesenchymal stem cells to be directionally differentiated into Leydig cells and application of the composition.

The invention provides a method for inducing umbilical cord mesenchymal stem cells to be differentiated into leydig cells through gold nanoparticle medicated gene transfection. Plasmids with SF-1 (seroidogenic factor 1) genes and LHR (luteotropic hormone receptor) genes are cotransfected into the umbilical cord mesenchymal stem cells through gold nanoparticles, then the umbilical cord mesenchymal stem cells are induced by compounds or growth factors to be directionally differentiated into the leydig cells. The leydig cell differentiation method is efficient and safe, the differentiated leydig cells can serve as a research substitution model and can be used as seed cells of clinical cell treatment of the leydig cells, and an application prospect of leydig cell regeneration replacement treatment is widened.

The invention relates to the field of cell cultivation and in particular relates to a separation and purification method of testicular interstitial cells of a naked mole rat. The interstitial cells of the naked mole rat are separated and purified by adopting enzymatic digestion and a Percoll continuous density gradient method. The method provided by the invention can obtain a lot of testicular interstitial cells of the naked mole rat with normal functional activity simply, efficiently and economically, and cultivation under a low oxygen condition can ensure that the cells still can keep the biological characteristics in an in-vivo state in an in-vitro environment, so that the special physiological functions of the testicular interstitial cells of the naked mole rat can be conveniently and directly researched in a pure in-vitro cell cultivation model further, thereby providing an important theoretical basis for exploring the biological mechanism and applying the same to the clinical related fields.

The invention provides a preparation method and application of a eucommia ulmoides male flower extract with an anti-fatigue effect. The preparation method comprises the following steps of: taking a certain amount of eucommia ulmoides male flowers, extracting the eucommia ulmoides male flowers with water of which the mass is 15 times that of the eucommia ulmoides male flowers, refluxing for 2 hoursat 100 DEG C, filtering and taking filtrate for later use, adding water of which the mass is 15 times that of the eucommia ulmoides male flowers into the eucommia ulmoides male flowers, boiling and refluxing for the second time, filtering and collecting filtrate, mixing the filtrate obtained in the two times, concentrating and performing spray drying to obtain the eucommia ulmoides male flower extract. According to the preparation method, the eucommia ulmoides male flowers are extracted with water, filtered and concentrated to obtain the extract; and experiments prove that the eucommia ulmoides male flower extract and monomer components thereof can promote secretion of testosterone in testicular interstitial cells, and can be used for health-care food with the main anti-fatigue function.

The invention discloses a method for testing influence of midazolam on development of testicular interstitial cells cultured in vitro. Comprising the following steps: culturing rats, separating CD90 positive cells, culturing testicular mesenchymal stem cells (SLC), detecting the influence of midazolam on proliferation and differentiation, amplifying the SLCs and detecting the cell activity, carrying out DAPI dyeing and counting, analyzing cells by a flow cytometer, analyzing testosterone (T) and progesterone (P4), carrying out mRNA and protein expression detection through QPCR (Quantitative Polymerase Chain Reaction) and WB (White Brown) experiments, and determining significant differences and drawing a chart through SNK (Sodium Nitrogen Kinase) detection. The steps are simple, and the influence of midazolam medicine on proliferation and differentiation of the testicular mesenchymal stem cells can be effectively verified.

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 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.