15 results about "Sertoli cells" patented technology

The invention discloses a method for generating haploid sperm-like cells through in-vitro differentiation of pluripotent stem cells and application of the haploid sperm-like cells. Human pluripotent stem cells are firstly differentiated into ectoderm-like cells, then the ectoderm-like cells are spontaneously gathered and converted into primordial germ cell-like cells under the induction of ectoderm induction factors, and then the primordial germ cell-like cells and testicular sertoli cells are subjected to mixed culture to form recombinant testicular tissues; the recombinant testis tissue enters meiosis under the induction of nutrition limitation and retinoic acid, then meiosis is completed under the stimulation of meiosis second-stage induction factors such as testosterone, follicle-stimulating hormone and pituitary extract, and interaction between primordial germ cell-like cells and testis supporting cells is enhanced by using small molecules SB431542, so that the meiosis of the testis is enhanced. And finally inducing to generate haploid sperm-like cells. According to the invention, a new tool can be provided for researching a molecular mechanism of male reproduction, and an important thought can be provided for research and transformation of asthenospermia, oligospermia, drug screening and the like.

This invention provides a biomarker for spermatogenesis disorders and its application, wherein the biomarker is the MNDA molecule. This application characterizes the functional abnormalities or spermatogenesis disorders of Sertoli cells by detecting the expression level of MNDA molecules on cells in testicular tissue sections, and can therefore be used for the clinical diagnosis of non-obstructive azoospermia.

The invention belongs to the technical field of medicines, and particularly discloses application of afzelin in preparation of an anti-testis aging medicament, and testis aging resistance is particularly improvement or recovery of an aged testis tissue structure. The invention also provides a medicament for resisting testis aging. The medicament contains afzelin. The invention provides a new application of afzelin, the afzelin can be used for saving senescence testicular tissue structure disorder, remarkably reducing the epididymal sperm deformation rate and increasing the number of supporting cells and undifferentiated spermatogonium cells, the new application value of afzelin is developed, and a theoretical basis is provided for developing afzelin into anti-reproductive aging drugs.

The invention discloses a reagent for editing a pig INSL3 gene and application of the reagent. The reagent for editing the pig INSL3 gene contains an INSL3-sgRNA A and an INSL3-sgRNA B, wherein the INSL3-sgRNA A and the INSL3-sgRNA The target sequence of the INSL3-sgRNA A is as shown in SEQ ID No.5; the target sequence of the INSL3-sgRNA B is shown in SEQ ID No. 6. Experiments prove that the INSL3-sgRNA A and the INSL3-sgRNA B are combined with a CRISPR (clustered regularly interspaced short palindromic repeats) / Cas9 system, so that the pig INSL3 gene can be efficiently edited, and the editing efficiency of the INSL3 gene of pig testicular cells and the editing efficiency of the INSL3 gene of pig fetal fibroblasts can reach 8.47% and 11.45% respectively. The invention has important significance on the function research of the INSL3 gene of pig cells or individuals and the breeding of pig genetic disease resistance.

The invention discloses a pilose antler active peptide and application thereof in anti-inflammation, anti-oxidation and anti-aging, and belongs to the technical field of biology. According to the invention, the brand-new active peptide SPLASDLDL screened out from pilose antler for the first time is utilized, and competitive inhibition on TNFR1 is realized through the characteristic that the brand-new active peptide SPLASDLDL is specifically combined with a TNFR1 receptor. A constructed sertoli cell senescence model proves that the polypeptide has anti-inflammatory, anti-oxidation and anti-senescence effects. The invention successfully solves the technical problem that in the prior art, the competitive inhibition TNFR1 targeting peptide cannot be found from pilose antler and the action mechanism of the TNFR1 targeting peptide cannot be clarified, and provides the action mechanism and scientific basis for developing novel anti-inflammatory and anti-aging drugs derived from natural products.

This invention discloses a method for the isolation, culture, and identification of chicken sexual gonadal cells, belonging to the field of biotechnology. The invention selects chicken embryos hatched to 18.5 days old, isolates the gonads after PCR sex identification, and uses 0.25% trypsin-EDTA digestion followed by filtration through a 70μm filter to obtain a single-cell suspension with a survival rate >90% and free from PGC contamination. A sex-specific culture system is established, allowing cells to be stably passaged and maintain a sex-specific phenotype. Through morphological, qRT-PCR, immunofluorescence, and Western blot multidimensional identification, accurate identification of male testicular Sertoli cells and female ovarian granulosa cells is achieved. This method is simple to operate, has good reproducibility, and produces high cell viability and purity. It can provide a stable gonadal microenvironment for the in vitro directed differentiation of chicken primordial germ cells, significantly improving gamete induction efficiency. It is suitable for research on avian reproductive and developmental mechanisms, transgenic breeding, and germplasm resource preservation.

The invention discloses an application of miR-486-3p targeting ACSL4 (acyltransferase) in regulation of testicular ferroptosis. The invention is based on the discovery that ACSL4 is a target gene of miR-486-3p in plateau zokor and participates in androgen regulated seasonal spermatogenesis. According to the invention, the functions of the miR-486-3p and the ACSL4 are researched by using a mouse. A dual-luciferase verification report shows that the expression of the miR-486-3p in a mouse is regulated and controlled by combining 3UTR of the ACSL4 gene. In vitro, testicular sertoli cells regulated by androgen signals are used as research objects, and the fact that the proliferation, oxidative stress and ferroptosis of the cells are regulated by miR-486-3p targeting ACSL4 is found. In vivo, oxidative stress and ferroptosis of mouse testis are caused by inhibition of miR-486-3p expression, and sperms are abnormal. MiR-486-3p is overexpressed in testis with ferroptosis, so that accumulation of peroxide in tissues is relieved, and sensitivity to ferroptosis is reduced.

The application discloses a method for extracting mitochondria-associated membranes (MAM) and other subcellular components from podocytes, which comprises the following steps: pretreating the podocytes, crushing the podocytes by low-power ultrasonic, separating the crude mitochondrial components by differential centrifugation, and then purifying the high-purity MAM and other subcellular components by density gradient centrifugation and ultracentrifugation. The method replaces the traditional homogenizer grinding with low-power ultrasonic crushing, improves the MAM extraction yield by controlling the ultrasonic parameters, appropriately simplifies the differential centrifugation steps, increases the yield while ensuring the purity, sequentially separates the unbroken cells, the crude mitochondria, the cytoplasm and the endoplasmic reticulum components, and finally obtains the high-purity MAM and other subcellular components by density gradient centrifugation and ultracentrifugation purification. The method effectively improves the yield of the podocyte separation and extraction of the MAM and other subcellular components, reduces the cost, and provides technical support for the research on the podocyte MAM proteomics.

The present invention relates to in vitro methods for production of Sertoli cells and related organoids. The Sertoli cells and testis like organoids may be used for therapeutic purposes including facilitation of the production of spermatogonia from pro-spermatogonia stem cells.

The invention relates to application of ginsenoside Rg4 as a unique active ingredient in preparation of a medicine for improving male delayed gonad hypofunction, belonging to the field of biological medicine. According to the invention, the action mechanism of the Rg4 for improving LOH is defined for the first time: the Rg4 can specifically up-regulate the mRNA expression quantity of StAR and StARD7 in a testis tissue, and promote the transport of cholesterol to the inner mitochondrial membrane of a testis Leydig cell from the transcriptional level; the Rg4 improves the mRNA expression quantity of P450scc, 3 beta-HSD in testicular tissues by promoting the efficient transport of cholesterol, enhances the conversion of cholesterol to pregnenolone and the catalytic synthesis of subsequent steroids, and improves the endogenous testosterone synthesis capability; the Rg4 can recover the negative feedback regulation function of an HPG axis, regulate secretion of pituitary LH and FSH to physiological steady state, and avoid abnormal fluctuation of hormone level. The invention fills the technical blank of Rg4 in the field of LOH gene regulation and control.

Provided is use of PGAM2 in the breeding of Blcattle black cattle, which belongs to the technical field of molecular biological breeding, in which individuals having a high PGAM2 content are screened as high-quality breeding bulls. In the present disclosure, it is proved by means of experiments that PGAM2 may promote the proliferation and glycolytic metabolism of Sertoli cells to produce lactic acid, an energy substrate required for spermatogenic cell development. It regulates the formation of blood-testis barrier and facilitates the communication between Sertoli cells and germ cells, thereby promoting spermatogenesis. The present disclosure provides a new direction for the breeding of Blcattle black cattle for high quality beef production.

This invention relates to the field of biomedical technology, and in particular to miR-143-3p and Prok2 Application of genes in regulating the proliferation or apoptosis of mouse testicular Sertoli cells. This invention demonstrates that overexpression of miR-143-3p significantly inhibits the proliferation of mouse testicular Sertoli cells and promotes their apoptosis, while inhibiting miR-143-3p expression promotes their proliferation and inhibits their apoptosis. Furthermore, this invention also found that reducing... Prok2 Gene expression levels can inhibit the proliferation of Sertoli cells in mouse testes and promote their apoptosis; increase... Prok2 The expression level of this gene can promote the proliferation of Sertoli cells in mouse testes and inhibit their apoptosis. Therefore, the miR-143-3p provided by this invention can be used to prepare products for the prevention and treatment of rodent infestations or for promoting rodent reproduction.

The invention provides a dyszoospermia biomarker and an application thereof. The biomarker is an MNDA molecule. According to the application, the state of dysfunction or dyszoospermia of Sertoli cells is represented by detecting the expression level of MNDA molecules on cells in a testicular tissue slice, so that the application can be used for clinical diagnosis and the like of non-obstructive azoospermia.

The invention belongs to the field of biological tissue engineering, and discloses a culture method for promoting testicular tissue structural maturation in vitro and application. The method comprises the following steps: cutting a testicular tissue into blocky fragments with the volume of 0.5-1 mm; a gas-liquid interface culture mode is adopted, the lower portion of a pre-balanced 1.5% sepharose gel cubic support is soaked in a basic culture medium, the upper portion of the pre-balanced 1.5% sepharose gel cubic support makes contact with air, and then tissue blocks are placed on the support; the method comprises the following steps: adding a WNT5A pathway agonist Foxy-5 with the final concentration of 200-400 [mu] M into a basic culture medium containing alpha MEM, 10% KSR and 1% P / S, and continuously culturing for 11 days under the conditions of 34 DEG C and 5% COs. According to the method, a WNT5A signal channel is continuously activated by using Foxy-5 through gas-liquid interface culture, so that the polarity of a supporting cell can be effectively maintained, and the integrity of a blood-testis barrier (BTB) and the morphology regularization of a seminiferous tubule are promoted. According to the invention, a standardized and repeatable testicular tissue in-vitro maturation platform is established, and the method can be used for research in fields related to reproductive toxicity evaluation, drug permeability test and spermatogenesis microenvironment.

This invention discloses a rapid method for detecting calcium signals in undifferentiated and differentiated spermatogonia. The method uses a mild enzymatic digestion method to prepare a testicular single-cell suspension. Indo-1 AM, Thy-APC, and c-kit-PE are used for staining and labeling. The cell population to be analyzed is determined based on the difference between the lateral and forward scattering areas of the cells in the testicular cell sample. Then, Thy-APC and c-kit-PE are used to label undifferentiated and differentiated spermatogonia, respectively, to further select specific cell groups. Finally, the fluorescence intensity changes of indo-1 AM in each specific cell subpopulation are detected, thereby obtaining the changes in calcium ion concentration in undifferentiated, undifferentiated-to-differentiated, and differentiated spermatogonia. This method can rapidly analyze the calcium ion concentration in undifferentiated, undifferentiated-to-differentiated, and differentiated spermatogonia. The method is simple and convenient, and provides a foundation for identifying spermatogonial dysfunction caused by calcium signal imbalance.