20 results about "Sertoli cell" patented technology

The invention discloses a porcine circular RNA, a porcine circLDHC, an overexpression vector and a construction method and application thereof, and relates to the technical field of livestock molecular biology. The porcine circular RNA is formed by cyclizing 3rd to 6th exons of an LDHC gene, and the nucleotide sequence of the porcine circular RNA is as shown in SEQ ID NO: 1. The porcine circLDHC is obtained by carrying out reverse transcription on porcine circular RNA (Ribonucleic Acid). According to the invention, a specific amplification primer is designed for circLDHC, and the real existence of circLDHC is verified through PCR (Polymerase Chain Reaction) amplification, sequencing and RNase R enzyme digestion. Fluorescent quantitative PCR analysis finds that circLDHC is highly expressed in sexual mature testis tissues of pigs, the expression level of circLDHC is related to testis maturation, and meanwhile, the expression quantity of circLDHC in testis of 75-day-old Meishan boars is remarkably higher than that of circLDHC in testis of same-day-old Duroc boars (plt; 0.01) of the substrate. The invention further finds that the over-expressed circLDHC inhibits the proliferation of the porcine sertoli cells, promotes the maturation of the porcine sertoli cells, lays a foundation for further researching the start of the puberty period of the boars, and provides a new thought for analyzing the sexual precocious puberty mechanism of the boars and improving the reproductive performance.

The invention discloses application of mangiferin in preparation of a protective agent for testicular sertoli cells and preparation of a medicine for treating male infertility diseases, and relates to the technical field of medicine application. Wherein the mangiferin can play a role in protecting the sertoli cells, and the protection role is reflected in ways of improving the cell viability of the sertoli cells, inhibiting the apoptosis of the sertoli cells, inhibiting the oxidative stress of the sertoli cells, inhibiting the inflammatory response of the sertoli cells and the like; meanwhile, mangiferin can be used for treating male infertility diseases in ways of relieving weight loss, repairing testis, improving endocrine system functions, inhibiting oxidative stress of testis tissues, inhibiting inflammatory response of the testis tissues and the like. Wherein the mangiferin can inhibit the inflammatory reaction mainly by inhibiting the activation of NLRP3 inflammasome to inhibit the NLRP3 pathway, so that the inflammation and even pyroptosis of testicular sertoli cells are remarkably reduced to improve spermatogenesis, and the male infertility disease is well treated.

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.

The invention discloses novel application of a GJB5 gene, a construction method of a GJB5 gene overexpression vector and a method for improving high temperature resistance of economic animals, and relates to the technical field of molecular biology. The preparation for overexpressing the GJB5 gene improves the heat resistance of the economic animal sertoli cells through overexpression of the GJB5 gene. Meanwhile, the invention also discloses a construction method of the GJB5 gene overexpression vector and a method for improving the high temperature resistance of economic animals. The effect of the GJB5 gene in heat stress is found for the first time, the gene is expected to be overexpressed in pigs or genetically modified into economic animals such as cattle and sheep in the future, so that the heat resistance of the pigs is improved, the semen quality of the pigs under the heat stress condition is further improved, and the GJB5 gene has important social benefits and economic benefits.

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 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.

The application provides application of an ACOT12 gene in regulation of proliferation of bovine testis Sertoli cells and belongs to the technical field of biotechnology.The application observes the proliferation of bovine testis Sertoli cells by interfering with ACOT12 gene expression.The results show that interference with the ACOT12 gene can significantly inhibit the proliferation of bovine testis Sertoli cells, which indicates that the ACOT12 gene has a positive regulation effect on the proliferation of bovine testis Sertoli cells.

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.

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 invention discloses novel application of a CLIC3 gene, a construction method of a CLIC3 gene overexpression vector and a method for improving high temperature resistance of economic animals, and relates to the technical field of molecular biology. The preparation for overexpressing the CLIC3 gene improves the heat resistance of the economic animal sertoli cells through overexpression of the CLIC3 gene. Meanwhile, the invention also discloses a construction method of the CLIC3 gene overexpression vector and a method for improving the high temperature resistance of economic animals. The effect of the CLIC3 gene in heat stress is found for the first time, the gene is expected to be overexpressed in pigs or genetically modified into economic animals such as cattle and sheep in the future so as to improve the heat resistance of the pigs and further improve the semen quality of the pigs under the condition of heat stress, and the CLIC3 gene has important social benefits and economic benefits.

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 discloses a yak sertoli cell separation method which comprises the steps of yak testis disinfection, yak testis tissue separation, cell digestion, cell purification and the like, so that the whole process of the yak sertoli cell separation method is realized. The separation method of the yak sertoli cells is simple and convenient to operate, the separated yak sertoli cells are relatively high in purity, other purification measures are not needed, the subsequent culture stability is good, and the separation efficiency is high; meanwhile, the aspects of cell pollution, separation purification and identification, separation method cost and the like can be effectively controlled, and popularization and application are facilitated.

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.

The present invention relates to a method for the in vitro production of Sertoli cells and related organoids. Sertoli cells and testis-like organoids can be used for therapeutic purposes, including promoting the production of spermatogonia from pre-spermatogonial stem cells.

The invention discloses a preparation method and application of a testis repair factor preparation, relates to the field of biological preparations, and aims to solve the problem that existing mesenchymal stem cells have a poor treatment effect on testis aging. The method comprises the following steps: 1, taking P1-generation umbilical cord mesenchymal stem cells, and culturing by adopting a special culture medium; 2, culturing the culture solution supernatant obtained after the culture in the step 1 in a Graviite microgravity simulation control system, then transferring the culture solution supernatant into an ultrafiltration tube, and centrifuging for ultrafiltration concentration to obtain a cell culture supernatant concentrated solution; and 3, adding normal saline into the cell culture supernatant concentrate obtained in the step 2 to prepare the testis repair factor preparation. The repair factor preparation prepared by the method has a good protection effect on testis intermediate cells and supporting cells, and can recover testosterone secretion and improve testis hypofunction by improving testis oxidative stress and immunoregulation.

The invention provides a separation method and an identification method of bactrian camel testis sertoli cells, and belongs to the technical field of cell separation. The method comprises the following steps: mixing 0.08-0.12% of collagenase IV solution and 0.23-0.27% of trypsin-EDTA (Ethylene Diamine Tetraacetic Acid) according to a volume ratio of 1: 1, digesting a bactrian camel testis tissue block, and then combining differential adhesion and hypotonic treatment to obtain bactrian camel testis supporting cells with the purity of 95% or above, meanwhile, reducing cell damage, and enabling the cell survival rate to exceed 85%; and the identification is completed by combining morphological observation, immunofluorescence staining and secretion function detection. The separated bactrian camel testicular sertoli cells can be used for double-hump reproductive performance research, and a foundation is laid for the reproductive performance of bactrian camels.

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.