79results about How to "Stable proliferation" patented technology

The invention disclose a technique for the tissue culturing and breeding of the red-welcome cuckoo as well as the technique for domestication and replanting, which comprises the first generation culture of inducing the clump sprout by applying the germ-free seedling as explant, subculture for enrichment-culture, root-growing inducing for the clump sprout, water-planting for domestication before replanting the tissue culture sprout, the final replanting to flowerpot and getting the red-welcome cuckoo. The invention is suitable for the red-welcome cuckoo from different places, the k-factor of the clump sprout is high, the root is uniform, and the survival rate after the water-culture domestication is high and is suitable for the red-welcome cuckoo commercial production.

The invention provides a NK (natural killer) cell culture medium and a culture method of a NK cell, wherein the NK (natural killer) cell culture medium comprises the following components in parts by weight: 83 to 93 parts of serum-free basal culture medium; 4 to 6 parts of plasma; 0.5 to 1.5 parts of interleukin-2; 0.5 to 1.5 parts of anti-CD3 monoclonal antibody; 1 to 5 parts of insulin-like growth factor 1; 1 to 3 parts of lycium barbarum polysaccharide. According to the NK (natural killer) cell culture medium provided by the invention, the safety is better, and the NK cell cultured by the culture medium with the components can quickly and stably proliferate, and has better killing activity to tumor cell. Experimental results show that, after using the NK cell culture medium provided by the invention for culturing the NK cell for two weeks, the proliferation times soars almost fiftyfold; the killing activity to K562 reaches 91 percent at effector-target ratio being 40: 1.

The invention discloses a combined algal reef cluster. Artificial algal reefs in the prior art only can provide attachments for growth of algae but can not make algae at part of sea areas be rapidly multiplied within short time. The combined algal reef cluster comprises floaters, slings, cultivating bodies and positioning anchors. Each cultivating body comprises a circular ring upper frame, a bottom plate, a middle column, a conical surface silica gel base body, a balancing weight and algal reef bricks, wherein the edge of the wide end of the conical structure of the conical surface silica gel base body is fixedly connected with the upper frame; the edge of the narrow end is fixedly connected with the edge of the bottom plate; the algal reef bricks are laid on the inner side surface of the silica gel base body; the top end of the middle column is connected with the corresponding floater through the corresponding sling; the bottom end of the middle column is connected with the corresponding positioning anchor through the corresponding sling. The combined algal reef cluster has the advantages that algal seedlings are protected, and the survival rate of algal seedlings is improved; the algal growing space is extended when algae grow to be a certain scale, and algae are further multiplied; the yield of a unit cultivating area is easy to control, and cultivating industrialization of algae is facilitated; algae automatically sink when big wind and waves come, and losses caused when algae are destroyed are reduced.

The invention provides a method for obtaining and cultivating sphagnum protonema. The method comprises the following steps: carrying out tissue smashing on sphagnum cormus used as a raw material; cultivating in a liquid culture medium; growing new protonema from fragments of sphagnum tissue cormus, and carrying out tissue smashing on the protonema; carrying out liquid cultivation, so as to obtain the sphagnum protonema. By adopting the method, a lot of protonemata of the sphagnum can be obtained; the cormus with uniform growth can be largely obtained by using the protonema within a short period of time; application of the sphagnum in a tissue culture physiological experiment and sphagnum protoplast preparation, transformation or regeneration is facilitated.

The invention provides a cell culture method for duck flavivirus. The method comprises the following steps: inoculating to attach an anchorage-dependent kidney cell BHK21 of a baby hamster to the surface of a cell culture container to form a monolayer; inoculating the monolayer with virus; culturing the monolayer inoculated with the virus in an incubator with 5% CO2 at 37 DEG C; and continuously passing by taking a cell culture product as an inoculum for the next passage. According to the cell culture method provided by the invention, after the virus is continuously passed for 5-7 generations on the monolayer, stable multiplication can be obtained; and after the cell is inoculated for 3-4 days, an obvious cytopathic effect can be formed. The method provides a convenient and visual virus operating platform for studying the biological characteristics of the virus and the molecular basis of virus pathopoiesia and for the vaccine research developed for effectively preventing and controlling the disease.

Cultivating a pluripotent stem cell in a medium comprising at least one member selected from the group consisting of ethanolamine, an ethanolamine analog, and a pharmaceutically acceptable salt thereof, and which is substantially free of β-mercaptoethanol or contains β-mercaptoethanol at a concentration of not more than 9 μM, and the like, is effective for the proliferation of a pluripotent stem cell while maintaining an undifferentiated state.

The invention refers to a process for using yeasting method to produce lactic acid. Takes out the mixed liquid of embedding agent and fungus suspending liquid to solidify the cells, the embedding agent is made up of fungus suspending liquid: 2% kala glue liquid: 8-12% polyvinyl alcohol and 2-4 sodium alginate with proportion 1:1:3-5, combines with using pH controller to control the acid rate pH value = 5.0-6.2 automatically, when the pH gauge displays the pH value is 5.0 in the yeasting liquid, the pH control system switches on the power automatically, the circular pump begin to work, when the pH value reaches 6.5, the relay cuts off the power, the circular pump stops, the yeasting is going on, it realizes in situ separation yeast lactic acid generating.

The invention provides an infectious bursal disease virus Vero cell-adapted strain and belongs to the field of bioengineering. The related infectious bursal disease virus Vero cell-adapted strain is named Ck /Jiangsu/ NJ-23/2008 and has a collection No. of CGMCCNO.8852. The infectious bursal disease virus Vero cell-adapted strain which can efficiently multiply on serum-free cultured Vero cell is finally obtained through wild strain separation, chick embryo passage, Vero cell passage adaption; the infectious bursal disease virus Vero cell-adapted strain is subjected to continuous passage culture on the serum-free cultured Vero cell and TCID50 can be kept to be higher than 108.5/mL. Virus culture solution is inactivated and prepared into oil emulsion; after the prepared oil emulsion is used to immunize chicken, detection proves that the prepared oil emulsion has good immunogenicity. The infectious bursal disease virus (IBDV) strain and the production process thereof are simple, safe and efficient and suitable for industrial culture.

The invention discloses an influenza virus vaccine strain. The invention discloses a virus. The amino acid sequence of each of PB2 protein, PB1 protein, PA protein, NP protein, NA protein, M1 protein, M2 protein, NS1 protein, NS2 protein and HA protein are respectively shown as follows: SEQ ID No. 26, SEQ ID No. 34, SEQ ID No. 33, SEQ ID No. 21, SEQ ID No. 29, SEQ ID No. 27, SEQ ID No. 28, SEQ ID No. 31, SEQ ID No. 32 and SEQ ID No. 16. The recombinant virus disclosed by the invention can be rapidly and stably proliferated in high yield. Compared with the method for preparing the vaccine by utilizing the chicken embryo, the recombinant virus has obvious advantage, and the inactivated vaccine of the recombinant virus can be used for protecting animals against the infection of influenza virus.

The invention provides a human placenta mesenchymal stem cell, a preparation method and an application thereof. The preparation method comprises the following steps: collecting, separating, culturing,cryopreserving, detecting, recovering, and the like. A high-purity high-activity mesenchymal stem cell can be acquired by only performing slide adherent culture after a human placenta chorion tissueblock is acquired in the separating process; the technological process is simplified; the cost of enzymic digestion is saved; the adherence and growth rate of primary cells are accelerated; the cell culture period is shortened; the introduction of more external interference factors is avoided, so that the process stability can be easily controlled. The multiplication capacity of the human placentamesenchymal stem cell acquired according to the invention is more stable than that of other mesenchymal stem cells; after the human placenta mesenchymal stem cell passes to P20 generation, the cell still can stably proliferate, and the cellular morphology, molecular surface antigen and adipogenesis osteogenesis differentiative potential thereof all meet the regulations for minimum standard of MSCidentification from international cell therapy association.

The invention discloses an establishment method for an immortal bone mesenchymal stem cell line of a Chang-Bai piglet. The method comprises the following steps: taking bone marrow cells of a Chang-Bai boar piglet, culturing the bone marrow cells with a nutrient solution for 20 to 28 h, then replacing a fresh nutrient solution, continuing culture and carrying out passage until F3-generation Chang-Bai piglet bMSCs with uniform cellular morphology is obtained; infecting the F3-generation Chang-Bai piglet bMSCs with a lentivirus liquid carrying a Large T gene and then carrying out cell screening and passage until a cell strain stably expressing Egfp is obtained; and subjecting the cell strain stably expressing Egfp to passage for more than 50 generations and carrying out cytobiological analysis and detection so as to obtain the immortal bone marrow mesenchymal stem cell line of the Chang-Bai piglet. According to the invention, a system capable of supporting stable culture of porcine bMSCs (bone mesenchymal stem cells) is obtained; rapid and stable propagation of the cells in vitro is promoted and biological characteristics of the original generation of the cells are maintained; it is proved that immortal bMSCs of the Chang-Bai piglet are obtained; and in-depth research on biological properties of the porcine bMSCs can be carried out and the bMSCs can be applied to research and development of drugs or vaccines.

The invention discloses an epinephelus lanceolatus head kidney cell line as well as a construction method and application thereof. The ELHK cell line is preserved in the Guangdong Microbial Culture Collection Center (GDMCC) on December 04, 2020, the address is the 5th floor of No.59 building, No.100, Middle Xianlie Road, Guangzhou City, Guangdong Province, the postal code is 510070, and the preservation number is GDMCC No: 61340. The Epinephelus lanceolatus head kidney cell line-ELHK cell line is obtained, the growth state is good, cell proliferation is stable, the cell morphology takes a fibroblast sample as the main morphology, continuous passage can be achieved (at present, the cells have already been transferred to more than 120 generations), ultralow-temperature cryopreservation and recovery can be achieved. The establishment of the cell line lays a foundation for related research of grouper germplasm resource preservation.