358 results about "In vitro incubation" patented technology

The invention provides an improved blastocyst culture solution as well as a preparation method and application thereof, and relates to the field of embryonic cell in-vitro culture. Based on a formula of a conventional blastocyst culture solution, colony stimulating factor-1 (CSF-1) and a transforming growth factor-alpha (TGF-alpha) are added, and the colony stimulating factor-1 and the transforming growth factor-alpha are dissolved in the conventional culture solution according to the concentration of 0.02-0.06 mmol / L and the concentration of 0.01-0.05 mmol / L; the obtained improved blastocyst culture solution can improve the mulberry embryo formation rate or blastocyst formation rate in in-vitro culture, promote the development of low-quality embryos and improve the mulberry embryo quality or blastocyst quality, and can be used for preparing medicines for promoting embryo implantation or subsequent growth and development.

The invention provides 3D tumor cell culture hydrogel as well as a preparation method and application thereof. The 3D tumor cell culture hydrogel is prepared from the following raw materials: polyvinylpyrrolidone and polyvinyl alcohol. The preparation method comprises the following steps: mixing a polyvinylpyrrolidone solution with a polyvinyl alcohol solution, adjusting the pH value, and freezing-unfreezing to obtain the 3D tumor cell culture hydrogel. The 3D tumor cell culture hydrogel provided by the invention does not need to be additionally added with a chemical cross-linking agent, and the material is high in biocompatibility and flexible in modulus adjustment, and can be well applied to a 3D in-vitro culture model of tumor cells.

The invention discloses application of a method for enhancing the treatment effect of salmonella bacteriophage, and relates to the technical field of microorganisms. According to the method, bacteriophage is added into salmonella for in-vitro culture, after continuous subculture, the salmonella is dropwise added to an SS plate containing the bacteriophage, and resistant bacteria of the bacteriophage are screened. The split bacteriophage is screened aiming at the resistant bacteria of the bacteriophage, and the split bacteriophage and the initial bacteriophage form a bacteriophage cocktail formula, so that the salmonellosis can be well prevented and treated, and the generation of the resistant bacteria is delayed. The method has the beneficial effects that compared with the existing method for screening the bacteriophage-resistant bacteria through a double-layer plate method, the method is more convenient, the operation is simple, and the screening efficiency of the bacteriophage-resistant bacteria is greatly improved. The bacteriophage cocktail contains the bacteriophage for cracking the resistant bacteria, so that the treatment effect of the bacteriophage is improved, and the practical application scene is quite high.

The invention discloses a preparation method of cells with LTo-like functions, which is characterized in that adipose tissue-derived stem cells are treated by LTo differentiation induction signal molecules to obtain cells with LTo-like phenotypes, secretion of cytokines and chemotactic factors of the cells is up-regulated, and the cells have recruitment and support capabilities on lymphocytes in vivo and in vitro. The problems that LTo cells are rare and difficult to obtain are solved, and the adipose-derived stem cells are higher in efficacy, easy to obtain and culture in vitro, capable of being massively amplified, low in cost, efficient and safe.

The invention discloses a rapid and efficient extraction method of extracellular vesicles rich in mitochondria. The extraction method comprises the following steps: (1) culturing mitochondrial donor cells in vitro; (2) treating the mitochondrial donor cells with cytochalasin D; (3) collecting the cells; (4) oscillating a cell suspension; according to the method disclosed by the invention, by combining cytochalasin D treatment with a subsequent gradient centrifugal separation step, a sample of extracellular vesicles rich in mitochondria can be efficiently extracted, meanwhile, the defects existing in a traditional mitochondrial transplantation technology are overcome, and a new thought and method are provided for clinical application of mitochondrial transplantation.

The invention relates to the technical field of in-vitro embryo culture, in particular to an in-vitro culture method and culture system of a mouse embryo. The method comprises the following steps: dropwise adding hydrogel onto a PDMS substrate, putting at least one, at least two, at least three, at least five or more embryos into hydrogel liquid drops, curing, and then adding a culture solution for embryo culture; wherein the Young modulus of the cured hydrogel liquid drops is 10 to 100 Pa. The in-vitro culture system comprises a PDMS (Polydimethylsiloxane) substrate and hydrogel liquid drops arranged on the PDMS substrate, and the hydrogel liquid drops are used for coating the mouse embryo; wherein the Young modulus of the cured hydrogel liquid drops is 10 to 100 Pa. When the method and the system are used for in-vitro embryo culture of mice, the embryos can normally develop and have a relatively high development rate within the in-vitro culture time of 9 days during in-vitro embryo culture.

It comprises a receptacle for the culture of plant material, a cap for closing an opening to said receptacle, and means for allowing the entry and / or exit of gas from said receptacle, and is characterized in that it comprises a container for receiving a liquid culture medium inside the receptacle, and support means for supporting the plant material on said reception container, wherein said container is attached to the lid of the receptacle in a position suitable for use, said lid including at least one medium inlet and / or exit conduit that connects with the container to be able to fill and / or empty said container through said conduit.

The invention discloses a three-dimensional biological printing meniscus model for simulating the mechanical heterogeneity of a natural meniscus and a preparation method of the three-dimensional biological printing meniscus model. The model comprises a structural PCL fiber frame (red region fiber circumferential arrangement and white region radial arrangement, and simulates the trend of natural collagen fiber), regional specific hydrogel (the red region contains GelMA, HAMA and SilMA, and the white region contains GelMA and HAMA), and partitioned cell components (the red region contains BMSC, HUVEC and CTGF, and the white region contains BMSC and TGF-beta3), the PCL frame and the hydrogel cooperate to form a mechanical gradient, and the compression modulus from the red region to the white region is simulated to continuously change. The preparation method comprises the following steps: designing a three-dimensional model, printing a PCL framework, preparing hydrogel and a cell suspension, preparing bio-ink, filling and printing by multiple nozzles, curing and culturing in vitro to obtain a target model. Compared with the prior art, the method has the advantages that the key problem of mechanical mismatch of an existing support is effectively solved, the model better fits a natural meniscus in mechanical property and biological function, and a more adaptive in-vitro model support is provided for individualized meniscus repair and transplantation substitution research.

The invention provides an immune cell culture medium and a culture method thereof, and the culture medium significantly improves the amplification efficiency and functional activity of immune cells by optimizing the combination of cytokines, small chemical molecules and bioactive polypeptides. According to the invention, cell factors such as IL-2, IL-7 and IL-15 are integrated, and a specific antibody and a specific peptide fragment derived from a tumor-associated antigen are combined, so that a culture environment is obviously optimized. The culture medium disclosed by the invention shows an excellent effect in amplification and function enhancement of various immune cells, and is suitable for in-vitro culture and function research. Particularly, a new technical support and a breakthrough scheme are provided for immune cell treatment in the aspects of enhancing the killing activity of immune cells and maintaining the phenotypic stability.

The utility model provides a cell culture box for cell in-vitro culture, which relates to the technical field of cell culture boxes and comprises a cell culture box, a control panel is arranged at the top of the cell culture box, a sealing door is rotatably connected to the outer surface of the cell culture box, a heating plate and a guide rail are fixedly mounted on the inner surface of the cell culture box, and the control panel is arranged on the control panel. According to the tray fixing device, through the arrangement of the wedge-shaped block, the fixing groove, the limiting rod and the first spring, when the tray moves on the guide rail, accurate positioning and stable clamping connection can be achieved, so that the tray is effectively fixed, and through the arrangement of the pull ring, the clamping block, the fixing frame and the rotating block, the tray can be effectively fixed. The unlocking or fixing of the tray can be easily realized during the operation of a user, and the operation convenience and safety of the device are improved.

The application provides a culture medium and a culture method of ovarian cancer primary cells. The culture medium comprises an MST1 / 2 kinase inhibitor, sodium pyruvate, forskolin, epidermal growth factor, gastrin, fibroblast growth factor 7, nicotinamide, SB431542, and fetal bovine serum. Compared with the existing culture method, the in-vitro culture using the culture medium of the application has higher amplification efficiency; the culture of the ovarian cancer primary cells using the culture medium can maintain the morphological structure and pathological characteristics of the primary tissue, and improve the success rate and survival rate of the ovarian cancer primary cell culture.

This invention discloses an antibody that specifically recognizes Toxoplasma gondii surface antigens and its applications. The invention utilizes hybridoma technology to establish a hybridoma cell line by fusing Balb / C mouse spleen cells with SP2 / 0 cells. Monoclonal antibodies were prepared from these cells using in vitro culture and in vivo ascites induction methods, yielding the monoclonal antibody SH12-S7. This invention provides clinical and research institutions with an antibody that specifically recognizes Toxoplasma gondii surface antigens. It can be used for ELISA assays to detect clinical samples, for immunohistochemical assays to diagnose pathological specimens, and provides a convenient research tool for toxoplasmosis research departments. It has broad application value in the diagnosis and detection of definitive host infections and provides technical support for epidemiological investigations and evaluation of control effects of Toxoplasma gondii.

The invention relates to a composite microbial inoculant and an evaluation method for the antibacterial effect thereof. The method optimizes the in-vitro culture mode of the microbial agent compounded by aerobic bacteria and anaerobic bacteria by simulating the intestinal environment of probiotics in an animal body, and adopts a facultative anaerobic culture method for the microbial compound agent, namely, the outer layer of a sealing film of a triangular flask is wrapped with four layers of preservative films, and the neck of the triangular flask is tightened by a rubber band. The facultative anaerobic environment of aerobic bacillus and facultative anaerobic enterococcus faecalis is considered in the microbial composite inoculant, in the subsequent oxford cup bacteriostasis evaluation experiment, the experiment result with the accurate bacteriostasis effect and good parallelism can be obtained, and the action effect of bacillus and enterococcus faecalis composite bacteria can be efficiently and scientifically evaluated.

The invention discloses construction of a Guangdong small-ear spotted pig spermatogonial stem cell in-vitro culture system and screening of surface markers, and belongs to the technical field of cell culture.The technical scheme includes that testis are broken firstly, then enzyme digestion, density gradient centrifugal purification and differential adherent purification are conducted, and purified spermatogonial stem cells are obtained; inoculating the obtained spermatogonial stem cells to feeder layer cells and culturing in a culture medium; the culture medium is a DMEM culture medium added with FBS and growth factors; the mass fraction of the FBS in the DMEM culture medium is 8-12%, an enzyme digestion method, a density gradient centrifugation method and a differential adhesion method are combined for application, the Guangdong small-ear spotted pig spermatogonia stem cells with certain purity are obtained, cells enriched and purified in vitro can proliferate massively, and an in-vitro culture system of the Guangdong small-ear spotted pig spermatogonia stem cells is established.

Production and maintenance of hematopoietic stem cells in in vitro culture systems has proven to be elusive. Disclosed herein are hematopoietic stem cell compositions that originate from organoids such as liver organoids. These organoids also comprise a rare type of immune cell that is not yet fully elucidated due to the difficulty in isolating said immune cell from biological samples. Also disclosed herein are methods of producing said hematopoietic stem cells and immune cells from organoids, as well as methods of expanding hematopoietic stem cells from other sources using these organoids.

The invention discloses a culture method of in-vitro uniform monolayer cells of prawns, and belongs to the technical field of biology. The method comprises the following steps: (1) obtaining lymphatic tissues of a last-stage prawn at the early stage of shelling, completely stripping an outer membrane to obtain internal tissues, and cutting the internal tissues into small tissue blocks; (2) cleaning the small tissue blocks, inoculating the small tissue blocks to a culture plate coated with a gelatin solution, adding a complete culture solution into an inoculation system at intervals within 12 hours after inoculation for culture, replacing the complete culture solution 24-48 hours after inoculation, and inducing tissue cells to continuously migrate to form an in-vitro cell layer; (3) blowing and beating the cell culture to obtain a cell suspension, and centrifuging to obtain a cell monolayer; and (4) suspending the cell monolayer by adopting a complete culture solution, then inoculating the cell monolayer into a culture plate coated with a gelatin solution, and culturing to obtain the in-vitro uniform monolayer cells of the prawns. According to the method, the prawn monolayer cells which are long in in-vitro survival time, uniform and stable and are cultured in vitro can be obtained, and an important tool is provided for related virology research.

The invention discloses a preparation method of a mouse skin organoid, which comprises the following steps: separating dermis and epidermis of an E18.5 mouse to obtain dermis cells, mixing the dermis cells with back skin cells of an E14.5 embryo mouse, mixing by using Matrigel to form cell spheres, and gradually forming the skin organoid in in-vitro culture; the organoid is mainly applied to batch production of organoid containing hair follicles. The invention relates to the technical field of stem cells and regenerative medicine. According to the preparation method of the mouse skin organoid, a cell source is improved in the experiment process, the mouse skin organoid containing hair follicles can still be formed after cells in the early development stage are added, and related developmental biology research is carried out. And the source of another dermal cell is not limited to late pregnancy, and also can be a newborn mouse, so that the source of seed cells is expanded, and the cost is reduced.

A microfluidic chip for in vitro culture, comprising: a chamber, the chamber comprising a primary channel and at least one side channel adjacent thereto, the chamber having a bottom surface and a top surface, the chamber being configured as an open-top chamber having at least one aperture extending through the top surface and in fluid communication with the primary channel; and a separator disposed between the primary channel and the at least one side channel, the separator extending from the bottom surface of the chamber toward the top surface of the chamber; and optionally, an insert, the insert having at least one projection configured obstruct access to the aperture when the projection is placed into register with the one aperture. A method, comprising: contacting a solid tumor cell residing in the primary channel of a microfluidic chip and a sample cell perfused to the primary channel.

The present invention relates to a method for selectively differentiating and separating neural crest cells containing human axial specialization information from pluripotent stem cells, and according to the method of the present invention, early neural crest cells corresponding to the embryonic neural tube formation stage and not having formed human axial information can be prepared. The neural crest cells with specific human body axial information can be prepared and subjected to specific separation and in-vitro culture, so that various neural crest cells can be obtained at a time, and the development process of tissues and organs can be revealed by researching the biological process of the neural crest cells with the axial information; thus, it is possible to develop various cell therapeutic agents optimized for body part characteristics.

The invention relates to an enhanced adipose-derived mesenchymal stem cell and application thereof in preparation of a medicine for treating enteritis, and belongs to the technical field of cell biology. The invention provides an enhanced adipose-derived mesenchymal stem cell. The enhanced adipose-derived mesenchymal stem cell is an adipose-derived mesenchymal stem cell of an overexpressed CNDP2 gene. According to the method, only a carrier for expressing the CNDP2 gene needs to be transfected in the adipose-derived mesenchymal stem cells, a complex and difficult-to-control gene editing technology is not needed, the operation is simple and easy, and the operation can be completed in a common laboratory, so that the aging process of the adipose-derived mesenchymal stem cells can be delayed, and the in-vitro culture activity and the in-vivo enteritis resistance of the adipose-derived mesenchymal stem cells can be enhanced. The experimental period is short, the use condition is not harsh, and clinical application can be better adapted.

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.

The present invention relates to a medium composition for in vitro fertilization and / or in vitro culture comprising a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof, and a method using same. The compound or composition suppresses reactive oxygen species and lipid peroxidation in ovarian granulosa cells to preventing cytotoxicity and mitochondrial dysfunction caused by apoptosis and ferroptosis, leading to an improvement in the quality of aged oocytes, and to increase the developmental rate and blastocyst formation rate of pre-implantation embryos, thereby improving the efficiency of in vitro fertilization. In addition, treatment with the medium composition during the vitrification process of in vitro–fertilized embryos improves re-expansion and survival rates, thereby reducing structural and metabolic damage. Accordingly, the present invention can be advantageously applied to an assisted reproductive technology for treating infertility and subfertility, and in vitro fertilization and / or in vitro culture for improving the propagation efficiency of livestock.

The invention provides a method for obtaining an extract of plant origin from an undifferentiated or dedifferentiated plant cell line by in vitro culture, the method comprising the addition of at least one aglycone flavonoid to the culture medium. The obtained extract contains a group of target secondary metabolites. It can be advantageously used for the non-therapeutic cosmetic treatment of the skin and appendages.

Described herein are devices for applying an alternating electric field to a living subject or an in vitro medium at a frequency between 100 kHz and 500 kHz. Also described herein are methods of using the described devices for applying an AC electric field to a target region comprising rapidly dividing cells, e.g., cells associated with a variety of disorders or conditions. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present disclosure.

The invention provides a coffee - plant cell culture composition comprising a) roasted and ground coffee and b) a composition comprising plant cell culture grown in vitro, optionally in a form of a powder. In some embodiments, the coffee - plant cell culture composition is in a pod or container and may be used in a coffee machine. In some embodiments, the plant cell culture grown in vitro is grape berry cell culture grown in vitro (RGC).

The invention discloses application of spermidine in in-vitro culture of human unfrozen oocytes and embryos. By adding the combination of spermidine, melatonin and astaxanthin into the thawed oocyte resuscitation culture solution and the embryo in-vitro culture solution, the mitochondrial autophagy and mitochondrial functions of the thawed oocytes are improved, so that the in-vitro culture effect of the thawed oocytes and embryos is improved, and the success rate of an assisted reproduction technology is increased.

The patent discloses an antibody against human IgG, a preparation method and application thereof, a heavy chain variable region of the antibody comprises heavy chain complementarity determining regions HCDR1, HCDR2 and HCDR3, and amino acid sequences are respectively shown as SEQ ID NO:1-3; a light chain variable region comprises light chain complementarity determining regions LCDR1, LCDR2 and LCDR3, and amino acid sequences are respectively shown as SEQ ID NO:4-6. The application efficiently prepares a high-activity monoclonal antibody which can combine with different subtypes of human IgG Fc by using an in-vitro culture method of B cells, and specifically relates to two monoclonal antibodies which can specifically combine with human IgG1 protein, and provides a powerful tool for detecting human IgG1 protein and researching diseases related to human IgG1 protein.

This invention discloses an in vitro maturation culture medium for ferret oocytes and its preparation method, belonging to the field of animal embryo engineering technology. It comprises the following components: TCM199 basal culture medium, 50 mL fetal bovine serum, 25 IU follicle-stimulating hormone, 25 IU luteinizing hormone, 0.5 mg 17β-estradiol, 0.005 mg epidermal growth factor, 12.1 mg sodium pyruvate, 0.2 mM cysteine, 0.1 mM cysteine, 0.030 g penicillin, 0.025 g streptomycin sulfate, as well as an antioxidant complex, a combination of growth factors, and extracellular matrix components. This invention provides a comprehensive and suitable nutritional environment for ferret oocytes by precisely formulating a composite culture medium containing multiple key components. Specifically, the hormone combination at specific concentrations accurately simulates the in vivo physiological environment, effectively regulating the oocyte maturation process; multiple growth factors work synergistically to promote oocyte growth, development, and differentiation; and the addition of the antioxidant complex effectively eliminates free radicals generated during in vitro culture, reducing oxidative stress damage to oocytes.

The application provides a kind of liposome that can enhance organ targeting, preparation method and application, belongs to the field of biological medicine and nanotechnology.The application first proposes the strategy of actively regulating LNP organ targeting by pre-assembling specific protein crown in vitro, which breaks through the technical limitations of traditional passive formation of natural protein crown.Through selecting main lipids (CDO, CDA, EDM) with different head chemical structures, precise control of protein crown composition is realized.Furthermore, the method of the application is simple in process and mild in condition, only needs standard in vitro incubation steps, and has great clinical transformation potential.The pre-assembled protein crown method provided by the application significantly enhances the organ targeting efficiency of LNP: after pre-incubation of LNP1 with APOA1, the enrichment degree of spleen is increased by 2.8 times;after pre-incubation of LNP2 with VTN, the enrichment degree of lung is increased by 52%;after pre-incubation of LNP3 with APOE, the enrichment degree of liver is increased by 1.2 times.

The invention discloses a construction method of a Klf6 gene knockout mouse embryo model, which comprises the following steps: S1, preparing a gene editing solution containing Cas9 enzyme and sgRNA for targeting a third exon region and a fourth exon region of a mouse Klf6 gene by utilizing a CRISPR / Cas9 gene editing technology; s2, taking out the fertilized eggs of the mouse, transfecting the fertilized eggs of the mouse with a gene editing solution, and then performing in-vitro culture on the fertilized eggs; s3, carrying out embryo transplantation on the two-cell stage embryo obtained in the step S2 into a pregnant female mouse, and then carrying out conventional feeding; and S4, 7-8 days after embryo transplantation, taking out the uterus and stripping the embryo to obtain the Klf6 gene knockout mouse embryo model. According to the method, gene knockout is carried out by using CRISPR / Cas9 in the fertilized egg period in combination with an electrotransfection method, and compared with an existing method, the construction process time can be greatly shortened, the working efficiency is obviously improved, and the success rate is high.