215 results about "Granular leucocyte" patented technology

The invention provides a strain of lactobacillus plantarum YS-AG23 with high antioxidant activity, and a rhizoma polygonati and radix angelicae sinensis fermented product obtained by fermenting rhizoma polygonati and radix angelicae sinensis with the strain of lactobacillus plantarum YS-AG23. In a D-galactose induced premature ovarian failure mouse model, the fermentation product can regulate up the levels of antioxidant and stress protection genes SOD2 and HO-1 and an anti-aging gene SIRT1 by regulating down the expression of a key gene AGER of an inflammation and oxidative stress pathway, so that the oxidative stress and inflammatory response of model mouse ovarian tissues are remarkably reduced; meanwhile, the serum hormone level can be specifically adjusted, the serum estradiol (E2) level can be increased, the follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels can be reduced to be close to the normal range, the expression of anti-mullerian hormone (AMH) and hormone regulatory gene LHR of ovarian reserve markers can be up-regulated, the proliferative activity and estrogen secretion function of follicular granulosa cells can be improved, and the effect of preventing ovarian reserve is achieved. The wet weight of atrophic uterus and ovary of the model mouse is remarkably increased, uterine and ovary atrophy is effectively reversed, and the ovarian reserve function of the D-galactose induced premature ovarian failure model mouse is recovered. Besides, in an H2O2-induced human dermal fibroblast aging model, the yeast can improve the activity of aging cells in a concentration-dependent manner and reduce the level of reactive oxygen species (ROS) in the cells; in addition, by up-regulating the expression of a collagen synthesis key gene COL1A1 and down-regulating the expression of a cell senescence marker gene p16, skin cell senescence caused by oxidative stress can be effectively improved. In conclusion, the polygonatum sibiricum and angelica sinensis leavening can synchronously improve premature ovarian failure and skin cell senescence caused by oxidative stress by accurately regulating and controlling oxidative stress, inflammatory response and tissue function related genes, and has wide application potential.

The invention discloses sturgeon ovary active peptide with ovary aging resistance and a preparation method of the sturgeon ovary active peptide. According to the invention, sturgeon ovary is taken as a raw material, and peptide with the highest ovarian aging resistance activity is selected from different enzymolysis combinations through an in-vitro anti-oxidation test and an ovarian granular cell and caenorhabditis elegans test; three sturgeon ovary active peptide sequences with ovary senescence resistance activity are screened by virtue of bioinformatics technologies such as mass spectrum identification, molecular docking and the like, and the amino acid sequences of the sturgeon ovary active peptide sequences are Phe-Met-Ile (FMI), Phe-Asp-Pro-Phe (FDPF) and Ser-Trp-Pro-Trp-Gln (SWPWQ). The sturgeon ovarian peptide prepared by the method not only has remarkable anti-premature ovarian failure activity and successfully solves the problem of high-valued utilization of sturgeon byproducts, but also can be used as a natural and efficient functional factor, is widely applied to functional products in the related fields of health care products, cosmetics, medicines and the like, and has a wide application prospect. And the method shows extremely wide market application prospects and huge economic values.

The invention discloses application of a Wnt2 gene in buffalo ovarian granular cells, and belongs to the technical field of cell engineering and gene engineering. Wnt2 is taken as a research object, the Wnt2 overexpression vector or Wnt2 siRNA is transfected to the ovarian granulosa cells respectively by constructing the Wnt2 overexpression vector and synthesizing small interfering RNA (Wnt2 siRNA), then changes of signal channel gene mRNA and protein level related to ovarian granulosa cell apoptosis, proliferation and E2 and P4 secretion are detected, and finally phenotypic changes of the ovarian granulosa cells are detected. Results show that Wnt2 can inhibit apoptosis of ovarian granular cells and promote proliferation and E2 and P4 secretion. By exploring the application of the Wnt2 in the ovarian granular cells, the application has good application in researching an ovarian follicle atresia mechanism, initial estrus initiation and the like.

The invention relates to application of an NR1D1 gene in regulation and control of granular cells, and belongs to the technical field of biology. The invention provides an application of an NR1D1 gene in regulation and control of granulosa cells, including an application of a sequence shown in siRNA of the NR1D1 gene in preparation of a product for regulation and control of sheep granulosa cells, and the sequence is shown in SEQ ID NO: 1 and SEQ ID NO: 2. According to the regulation method and product of the granulosa cells, the proliferation apoptosis and activity of the granulosa cells can be effectively regulated, and the expression of follicular development related genes can be regulated. The method has the beneficial effects that in-vivo or in-vitro regulation and control effects on follicles and / or ovaries are realized, and a method and a product guarantee are provided for research on development and physiological and biochemical functions of granular cells, follicles and ovaries.

The invention provides an establishment method of umbilical cord mesenchymal stem cells with high expression of IGF-1 (Insulin-like Growth Factor 1) and application of the umbilical cord mesenchymal stem cells in premature ovarian failure. The construction method of the umbilical cord mesenchymal stem cells with high expression of IGF-1 comprises the step of transfecting the umbilical cord mesenchymal stem cells with the IGF-1 gene, so that the umbilical cord mesenchymal stem cells highly express IGF-1. The invention also provides application of the umbilical cord mesenchymal stem cell with high expression of IGF-1 in preparation of drugs for treating premature ovarian failure. According to the invention, the IGF-1 gene is transfected to the hUC-MSC, so that the hUC-MSC highly expresses the IGF-1, and the hUC-MSC secretes a large amount of IGF-1. The ovarian function is improved through the immunoregulation effect of hUC-MS, and meanwhile, the effect of resisting apoptosis of granular cells is achieved through IGF-1 secreted by hUC-MS. And the treatment effect on the premature ovarian failure is achieved through synergism of the two effects.

The invention relates to the technical field of biological breeding, in particular to application of an RBX1 gene in cow ovarian granular cells. According to the invention, the RBX1 gene is specifically regulated on the ovarian granulosa cells, and the RBX1 gene is found to have the effect of promoting the proliferation of the granulosa cells; and the gene is provided for the creation of a new high-fecundity breeding material for gene editing cattle in the later period.

The invention discloses application of MY-1B in preparation of medicines for improving ovarian aging, belongs to the technical field of biological medicines, and aims to solve the problems of how to effectively delay ovarian aging and improve ovarian functions. Experiments prove that the mRNA m5C modification level of ovarian granular cells in the ovarian aging process is increased, and the expression level of methylated transferase Nsun2 is increased. Ovarian aging caused by mRNA m5C modification level increase can be improved by supplementing the MY-1B, which is embodied in that the MY-1B can reduce the mRNA m5C modification level of aging nest granular cells; the MY-1B can be used for effectively correcting the orbital cycle disorder of the aged ovary and delaying the decline of the ovarian function; the MY-1B plays an important role in reducing the apoptosis level of granulosa cells in senescent ovaries; the MY-1B can improve the storage capacity of ovaries and enhance the quality of oocytes, so that the fertilization rate and the embryonic development potential of the oocytes are improved.

The invention belongs to the technical field of biomedicine, and particularly relates to an alpha-cyperone bionic nano preparation as well as a preparation method and application thereof, the alpha-cyperone bionic nano preparation is prepared from a polylactic acid-glycolic acid copolymer, alpha-cyperone, a bionic membrane and follicle-stimulating hormone polypeptide, the biomimetic membrane is characterized in that alpha-cyperone is loaded on a polylactic acid-glycolic acid copolymer, the biomimetic membrane is biologically camouflaged on the outer layer, then the biomimetic membrane is modified by follicle-stimulating hormone polypeptide, and the amino acid sequence of the follicle-stimulating hormone polypeptide is as shown in SEQ ID NO.1. According to the alpha-cyperone bionic nano preparation disclosed by the invention, the alpha-cyperone is loaded by utilizing the polylactic acid-glycolic acid copolymer, so that the alpha-cyperone is easily dissolved in water, and the blood circulation half-life period is prolonged and the targeting capability of an ovary part is improved through biological camouflage of the bionic membrane; the modification of the follicle-stimulating hormone polypeptide further improves the targeting ability to granulosa cells, and liver and kidney toxicity and muscle injury are not found in the experiment process.

The invention relates to the technical field of molecular biology, and particularly discloses application of miR-615 in ovarian granular cells. The miR-615 is applied to any one of the following items: preparing a product for promoting the activity and / or proliferation of ovarian granular cells; preparing a product for promoting generation of estradiol and / or progesterone; preparing a product for promoting the expression of the 3 beta-HSD and / or StAR gene; preparing a product for inhibiting ACSL4 gene expression; and preparing a product competitively combined with the lnc-GAM. According to the invention, porcine ovarian granular cells are used as a material, the expression level of lnc-GAM-miR-615 / ACSL4 regulatory axis is up-regulated or down-regulated in the porcine ovarian granular cells by using gene overexpression and interference technologies, and the expression of the lnc-GAM-miR-615 / ACSL4 regulatory axis is determined by detecting cell proliferation activity, hormone secretion level and apoptosis gene expression in combination with a dual-luciferase reporter gene test. And the regulation effect of lnc-GAM mediation on the apoptosis of the porcine ovarian granular cells is researched.

The invention discloses a traditional Chinese medicine composition for repairing and / or activating residual ovarian functions of an aging ovary, and a preparation method and application thereof, and relates to the technical field of biology. The traditional Chinese medicine composition is prepared from the following components: radix astragali, radix angelicae sinensis, radix ophiopogonis, radix polygonati officinalis, semen cuscutae, radix morindae officinalis, fructus rubi, rhizoma dioscoreae and folium mori. The invention further provides application of the traditional Chinese medicine composition in repairing and / or activating the residual ovarian function of the senescent ovary and improving the quality of the senescent oocyte, the traditional Chinese medicine composition can increase the volume and weight of the senescent ovary, increase the number of follicles of all levels in the senescent ovary, reduce apoptosis of granular cells in the ovary and improve the survival rate of the senescent ovary. The abnormal rate of oocytes, oxidative stress of the oocytes and DNA damage are reduced, development and maturation of residual follicles are promoted and maintained, and the traditional Chinese medicine composition has a wide application prospect.

The invention discloses application of an SFRP1 gene in buffalo ovarian granular cells, and belongs to the technical field of cell engineering and gene engineering. According to the invention, SFRP1 is taken as a research object, an SFRP1 overexpression vector or siRNA-SFRP1 is respectively transfected to ovarian granular cells by constructing the SFRP1 overexpression vector and synthesizing small interfering RNA (siRNA-SFRP1), then changes of signal channel gene mRNA and protein level related to ovarian granular cell apoptosis, proliferation and E2 and P4 secretion are detected, and finally phenotypic changes of the ovarian granular cells are detected. Results show that the SFRP1 can inhibit apoptosis of ovarian granular cells and promote proliferation and E2 and P4 secretion. The application of the SFRP1 in the ovarian granular cells is explored, and the SFRP1 has good application in research of an ovarian follicle atresia mechanism, initial estrus initiation and the like.

The invention discloses an MMP2 gene promoter region SNP (Single Nucleotide Polymorphism) site and application thereof in pig breeding. According to the invention, an MMP2 gene promoter region is taken as a research object, and the relevance between the MMP2 gene promoter region and sow sexual maturity is researched by adopting a molecular and cell biology method, so that the situation that g.30088284G > A site and g.30088004C > T site are obviously related to the group maturity day age of Dumet black pigs is found; then different vectors are transfected, and different influences of different genotypes of the sites on the activity of the IRS1 gene promoter are studied. Further, sow ovarian granular cells are used as experimental materials, experimental technologies such as transcriptome sequencing, a total iron colorimetric method and a malondialdehyde colorimetric method are adopted, the influence of the MMP2 gene on the transcriptome level and ferroptosis of the sow ovarian granular cells is researched, and it is found that cell transcriptome differential genes influenced by the MMP2 gene are mainly enriched to signal channels such as cell differentiation; and ferroptosis of cells is promoted.

The invention provides an ovary-targeting ROS response type SS31 nano-particle and a preparation method and application thereof, and belongs to the field of biomedicine.The ovary-targeting ROS response type SS31 nano-particle is constructed by utilizing a porous metal organic framework to carry hyaluronic acid and utilizing an ROS response type thioketone linker to be combined with mitochondrial targeting peptide SS31, and the ovary-targeting ROS response type SS31 nano-particle is used for preparing the ovary-targeting ROS response type SS31 nano-particle. And the SS31 is targeted and delivered to the ovarian tissue. The nanoparticles target ovary and slowly release SS31 under the action of ROS, so that oxidative stress of oocytes and granular cells caused by female reproductive system diseases such as premature ovarian failure, polycystic ovarian syndrome and the like is reduced, and the mitochondrial function of the oocytes and the granular cells is improved; and a foundation is laid for the subsequent clinical treatment of diseases related to the mitochondrial dysfunction of the female reproductive system by using the SS31.

The invention relates to the technical field of ovarian granule cell apoptosis, and discloses a method for regulating oxidative stress induced sheep ovarian granule cell apoptosis by using allicin, and the method comprises the following steps: firstly preparing a culture solution, then preparing an allicin stock solution, and identifying sheep ovarian granule cells; the method comprises the following steps: firstly, measuring the activity of ovarian granular cells after independent treatment of H2O2 and allicin, determining the optimal treatment time and concentration of H2O2 and allicin, then carrying out co-treatment of H2O2 and allicin, detecting the apoptosis of the granular cells by using flow cytometry, detecting the expression of apoptosis-related genes by using real-time fluorescent quantitative PCR, detecting the secretion of steroid hormones by using ELISA, and carrying out data analysis. The invention solves the problems that in the prior art, an effective regulation and control method for ovarian granular cell apoptosis induced by oxidative stress is lacked, and traditional medicines possibly have large side effects, unobvious effects and the like.

The invention provides an application of a mesenchymal stem cell modified by an IGF-1 (Insulin-like Growth Factor 1) gene in treatment of premature ovarian failure. The invention provides a modified IGF-1mRNA (Insulin-like Growth Factor). The modified IGF-1mRNA comprises a 5'end capping structure (Cap) and a 3 '-polyA tailed structure of an RNA (Ribonucleic Acid) sequence, and encapsulating the modified IGF-1 mRNA (messenger Ribonucleic Acid) by using an ionizable lipid. The invention further provides a preparation method of the liposome based on the ionizable lipid, a method for improving the transfection efficiency of the IGF-1 in-vitro transfected mesenchymal stem cells and the IGF-1 transfected mesenchymal stem cells obtained by adopting the method, MSCs are promoted to continuously and stably express IGF-1, the MSCs can stably secrete factors to promote cell proliferation and differentiation, and the granular cell secretion activity is maintained. The IGF-1 transfected mesenchymal stem cells can be used for preparing a medicine for treating premature ovarian failure, the IGF-1 modified MSCs can assist the premature ovarian failure in recovery, and the technical problem that the treatment effect cannot be improved due to the fact that the dosage of the MSCs is limited in the clinical treatment process can be solved.

The invention provides an application of a CTH inhibitor and / or an xCT inhibitor in preparation of a medicine for inhibiting mammal embryo implantation, and relates to the technical field of medicines. The application effect of a cystathionine-gamma-lyase (CTH) inhibitor and / or a solute carrier family 7 member 11 (xCT) inhibitor in inhibition of mammalian embryo implantation is verified through research. The research result shows that the xCT inhibitor and / or the CTH inhibitor can obviously inhibit proliferation of corpus luteinized granular cells and secretion of progesterone of a model mouse; meanwhile, by injecting the xCT inhibitor and / or the CTH inhibitor into the mouse model in vivo, ovary ovulation and the maturation rate of the ovulated oocyte nucleus can be remarkably inhibited, functional corpus luteum formation and embryo implantation can be remarkably reduced, and then a new thought and method are provided for development of contraceptive drugs.

The invention discloses application of spermidine in preparation of an agent for resisting mitochondrial oxidative damage and copper death damage, and relates to the technical field of biology. The invention provides an application of spermidine in preparing an agent for resisting mitochondrial oxidative damage and copper death damage, and further provides an application of spermidine in inhibiting copper death induced by copper sulfate and relieving iron-sulfur cluster protein loss and ester acylation protein aggregation induced by copper accumulation. The spermidine can obviously relieve cell viability decline induced by copper sulfate, reverse ROS and MDA level increase and mitochondrial membrane potential decline induced by copper sulfate and relieve iron-sulfur cluster protein loss and ester acylation protein aggregation induced by copper accumulation, and has obvious effects of inhibiting mitochondrial oxidative damage and copper death. The invention provides a method for effectively relieving copper death of ovarian granular cells, and solves the problem that the existing antioxidant and cytoprotective agent for resisting mitochondrial oxidative damage generate side effects in the use process.

The invention discloses application of amniotic mesenchymal stem cells in preparation of a medicine for improving and / or treating premature ovarian failure. The invention finds that the amniotic mesenchymal stem cells can significantly improve the granular cell proliferation ability and hormone secretion ability, and the premature ovarian failure animal model proves that the amniotic mesenchymal stem cells have leading treatment effects in the aspects of restoring the ovarian functions (hormone secretion, ovary volume and follicle number of each level) of POF mice and improving fertility again. And a foundation is laid for optimizing a treatment strategy of premature ovarian failure and promoting clinical application of mesenchymal stem cell treatment.

The present invention discloses the application of MY-1B in the preparation of drugs for improving ovarian senescence, belonging to the technical field of biomedicine, aiming to solve the problem of how to effectively delay ovarian senescence and improve ovarian function. Experiments have proved that during ovarian senescence, the mRNA m<supgt;5< / supgt;C modification level and the expression level of methyltransferase in ovarian granulosa cells increase. Supplementation with MY-1B can improve the condition caused by mRNA m Nsun2 expression.

The invention belongs to the technical field of biological medicine and livestock breeding, and particularly relates to application of anthocyanin in improving the reproductive function of female beef cattle by adjusting intestinal flora and reducing follicular granule cell apoptosis. Specifically, the invention discovers and verifies for the first time through research that the anthocyanin added into the feed can play roles in regulating the reproductive hormone level of the cattle with the ovarian dysfunction, improving the ovarian function and improving the intestinal flora lineage, and meanwhile, the anthocyanin can resist the apoptosis of ovarian granular cells by resisting oxidative stress injury, so that the reproductive function of the cattle is improved. The invention provides a scientific basis for the application of the anthocyanin in the aspect of improving the reproductive capacity of the female beef cattle, and discloses a mechanism for improving the ovarian function by resisting oxidation and regulating intestinal flora, thereby being beneficial to improving the application prospect of the anthocyanin in animal husbandry, and having a good practical application value.

The invention discloses a preparation method of mesenchymal stem cells for resisting ovarian granular cell aging. The preparation method comprises the following steps: providing umbilical cord blood stem cells; carrying out induced culture on the umbilical cord blood stem cells, collecting a supernatant after the culture is completed, and separating the supernatant to obtain an exosome; providing mesenchymal stem cells, and performing induced culture on the mesenchymal stem cells by adding the exosome, so as to obtain the required mesenchymal stem cells for resisting ovarian granular cell senescence. According to the preparation method of the mesenchymal stem cells for resisting ovarian granular cell senescence, the mesenchymal stem cells induced by the umbilical cord blood stem cell exosome can be prepared, and the mesenchymal stem cells can be used for treating premature ovarian failure in a whole body vein use mode. The umbilical cord mesenchymal stem cells induced by the stem cell exosome can exert a better immune regulation function, regulate the balance of an immune system and reduce immune-related inflammation and injury so as to delay the senescence of ovarian granular cells.

Compositions useful for delaying reproductive aging in an individual comprise L-ergothioneine and ginsenoside Rb1. Methods of delaying reproductive aging, such as methods of treating age-related ovarian granulosa cell injury.

The invention discloses an antioxidant nano material as well as a preparation method and application thereof, and belongs to the technical field of material preparation. On the basis of the reduction reaction of GSH and selenium-containing substances, oxidation of selenium is effectively protected by utilizing a protective atmosphere, and it is ensured that the selenium element is combined with GSH in a stable valence state. Finally, a novel selenium nano material (GSH-Se) is synthesized, and the GSH-Se has antioxidant ability and GSH-Px-like activity, can relieve oxidative stress induced primary yak follicular granular cell and mouse ovary steroid hormone synthesis disorder, cell apoptosis and mitochondrial division fusion abnormality, and can be used for preparing a novel selenium nano material (GSH-Se). Meanwhile, the GSH-Se can regulate and control steroid hormone synthesis and cell apoptosis of the yak follicular granular cells by inducing mitochondrial fusion, and a new thought is provided for application of the GSH-Se to improvement of female reproductive performance.

The invention belongs to the technical field of biological medicine, and particularly relates to a reagent for improving ovarian development and a preparation method and application thereof. The reagent for improving ovarian development is prepared by mixing the following raw materials in parts by weight: 20-30 parts of N-acetylcysteine; 5 to 10 parts of tranexamic acid; 3 to 8 parts of creatine; and 3000 parts of water. According to the reagent, the mRNA expression of PKD1P6 and follicle stimulating hormone receptors (FSHR) in ovarian tissues is up-regulated, the activity and functions of cumulus granular cells are enhanced, and apoptosis of the cumulus granular cells is inhibited, so that the effects of effectively promoting follicle development and inhibiting the apoptosis of the cumulus granular cells are achieved.

The invention relates to the technical field of animal husbandry, in particular to a synchronous oestrus-timing insemination method suitable for cattle, which comprises the following steps: firstly, performing intramuscular injection of PG (cloprostenol sodium) at different time points after calving to promote uterine recovery and luteolysis and lay a synchronous oestrus foundation; the method comprises the key steps that GnRH (gonadotropin release hormone) is given through intramuscular injection on the 43rd day, the 53rd day, the 60th day and the 69th day, pituitary is promoted to release follicle-stimulating hormone (FSH) and luteinizing hormone (LH), collection and screening of follicles are guided, follicle maturation is promoted, the oestrus uncovering rate and the oestrus conception rate are increased, a melatonin implant is implanted in a subcutaneous implantation mode on the 63rd day, and the melatonin implant is implanted in a subcutaneous implantation mode on the 63rd day. The synthesis of ovarian hormones is promoted by regulating the hypothalamus-pituitary-gonad axis, the secretion of follicle cell factors and maturation promoting factors is promoted, the development of granular cells is promoted, and the maturation of oocytes is promoted. According to the method, the pregnancy rate of a double-synchronization-timing insemination scheme which is most widely used in a large-scale dairy cow breeding pasture at present can be remarkably increased, and feasible technical support is provided for success of increasing the breeding efficiency of cows.

This invention provides the application of CTH inhibitors and / or xCT inhibitors in the preparation of drugs for inhibiting mammalian embryo implantation, relating to the field of pharmaceutical technology. This invention verifies the efficacy of cystathionine-γ-lyase (CTH) inhibitors and / or solute carrier family 7 member 11 (xCT) inhibitors in inhibiting mammalian embryo implantation. The results show that xCT inhibitors and / or CTH inhibitors can significantly inhibit the proliferation of luteinized granulosa cells and progesterone secretion in mouse models. Furthermore, in vivo injection of xCT inhibitors and / or CTH inhibitors into mouse models not only significantly inhibits ovulation and the maturation rate of released oocytes, but also significantly reduces the formation of functional corpus luteum and embryo implantation. Therefore, this application provides new ideas and methods for the development of contraceptive drugs.

The application discloses application of NCOA7 circular RNA nanoliposome in preparation of a drug for treating premature ovarian failure or ovarian reserve function decline, and belongs to the technical field of biological medicine.The present application proves that nuclear receptor coactivator 7 (NCOA7) can relieve ovarian aging and significantly improve the ovarian function and fertility of DOR patients.The present application finds that NCOA7 circular RNA nanoliposome treatment can significantly improve the NCOA7 protein expression level of DOR female ovarian granulosa cells, improve the aging phenotype and growth rate of the DOR female ovarian granulosa cells, and has no significant influence on the biological characteristics of the ovarian granulosa cells, thereby providing a new scheme for rescuing the ovarian function and improving the fertility of DOR women.

The invention provides a novel application of a raspberry extract. It is found through tests that any one of isoquercitrin, ellagic acid, vanillic acid, oleanolic acid, kaempferol-3-O-rutinoside, kaempferol-3-O-sophoroside, tiliroside, tormentic acid and raspberry extract can remarkably improve the activity of the human ovarian granular cells or remarkably improve the oxidative stress level, and the activity of the human ovarian granular cells can be remarkably improved. According to the present invention, the ovarian function can be effectively protected by the eight components, the ovarian function can be effectively protected by the eight components, or the mitochondrial membrane potential level can be significantly regulated and controlled, or the estrogen level in the microenvironment can be effectively maintained, such that the ovarian function can be effectively protected by the eight components, and the raspberry extract containing the eight components. The invention provides a new choice for ovarian function protection products, and also provides a new application for the substances.

The primary culture method of the porcine ovary granular cells comprises the following steps: step 1, taking a fresh porcine ovary, quickly putting the porcine ovary into a Hank's balanced salt solution preheated at 38 DEG C for transportation, repeatedly washing the ovary with the Hank's balanced salt solution to remove blood and impurities, and then extracting follicular fluid with a 5ml syringe under a sterile condition; according to the method, the granular cells can be effectively separated and cultured from the porcine ovary, and the porcine ovary granular cells with higher purity and better activity can be obtained, so that the subsequent cell biology experiment research, such as gene transfection, protein expression analysis and the like, is facilitated; a cell model is provided for researching the physiological functions (such as hormone secretion and follicle development regulation and control) of the porcine ovarian granular cells, the operation is relatively simple, the cost is relatively low, and the used reagents and instruments are common in a cell culture laboratory and are convenient to popularize and apply.

The invention relates to an isolated culture method of functional Chinese softshell turtle ovarian granular cells. The isolated culture method comprises the following steps: S1, collecting ovarian tissues: collecting Chinese softshell turtle ovarian tissues; s2, separating and collecting a granular cell layer; s3, digesting and collecting granular cells; s4, primary culture of granulosa cells: transferring the collected cells into a cell culture bottle containing 4 ml of a complete culture medium for primary culture, wherein the complete culture medium comprises the following main components: a DMEM culture medium, 15% of fetal calf serum, 1% of penicillin streptomycin and 1% of an insulin-transferrin-selenium additive; and S5, subculturing the granular cells. The isolated culture method and the culture medium system provided by the invention are simpler and more time-saving and labor-saving; according to the method, the aromatase CYP19A1 can be expressed normally, the characteristics of the granular cells are kept, the aromatase CYP19A1 can be expressed normally, it is indicated that the in-vitro cultured cells can keep the functionality of the cells, and the in-vitro proliferation capacity of the granular cells is improved to a great extent.