106results about How to "Improve the success rate of cultivation" patented technology

The invention discloses a culture medium for culturing fruiting bodies of Antrodia cinnamomea and a cultural method thereof. The culture medium comprises potato dextrose broth (PDB) with a concentration of 1-2%(w / v), glucose with a concentration of 2.8-3.6%(w / v), agar with a concentration of 2.2-4%(w / v) and a Chinese medicine composition with a concentration of 16.7-4.%(w / v). The cultural method comprises the following steps: preparing a solid culture medium according to the ratio of the ingredients, inoculating an Antrodia cinnamomea strain to the solid culture medium, putting the solid medium containing the Antrodia cinnamomea strain in a specific environment to culture for a certain period to generate the fruiting bodies of Antrodia cinnamomea, wherein the fruiting bodies of Antrodia cinnamomea obtained by the cultural method have same index components as wild Antrodia cinnamomea.

Provided is a plant tissue culture bacteriostatic (antiseptic) agent containing botanical fungicide. The bacteriostatic (antiseptic) agent comprises isothiazolinone, nisin and aqueous extracts of the Chinese herbal medicines of terminalia, groundsel and aromatic madder. When the bacteriostatic (antiseptic) agent is applied in plant tissue culture, 0.35 to 0.5% of the bacteriostatic (antiseptic) agent is added into a medium, which enables fungal and bacterial contamination to a culture to be effectively reduced or prevented and poses little adverse influence on plant growth, thereby enhancing the success rate of plant tissue culture; the bacteriostatic (antiseptic) agent can also be applied in open type plant culture, e.g., in the rooting phase of sound seedlings during a late plant culture stage, the bacteriostatic (antiseptic) agent is directly added into a medium, and the medium can be inoculated in indoor environment with bacteria or a few bacteria without undergoing the step of autoclaving or operating on a superclean bench, thereby simplifying seedling culture in plant tissue culture, enhancing working efficiency, saving a great amount of energy and reducing production cost for plant tissue culture seedlings; therefore, the bacteriostatic (antiseptic) agent has a critical actual application value.

A watermelon seed treatment method being high in germination rate comprises following steps: (1) pre-treatment of seeds: soaking watermelon seeds in an acidic soaking solution with flat seeds rejected; (2) preparation of a seed soaking liquid: mixing ammonium nitrate, sodium phthalate, sodium humate, an nutrient additive liquid and gibberellins to obtain a mixed aqueous solution; (3) seed soaking: soaking the pre-treated seeds in the seed soaking liquid; (4) sterilization treatment: treating the treated seeds under an infrared light lamp and sowing the seeds after sterilization. In the invention, the seeds are subjected to the pre-treatment so that dormant period of the watermelon seeds is broken, thereby promoting germination of the watermelon seeds. The seed soaking liquid is rich in nutrients so that the watermelon seeds can go through the dormant period with a short time, so that the speed of germination and seedling of the watermelon seeds is increased. The irradiation of the infrared light achieves the sterilization effect and can enhance disease resistance of the seeds, thereby increasing the cultivation successful rate of watermelon and achieving a high germination rate.

The invention belongs to the technical field of modern biotechnology cell culture. A purpose of the invention is to provide a tumor stem cell suspension culture method, which can be used for culture of tumor cells with different tissue sources, and tumor stem cells obtained through the culture method have characteristics of self-renewal capacity and enrichment. The tumor stem cell suspension culture method comprises: carrying out digestion centrifugation collection on tumor cells requiring culture, adopting PBS to carry out resuspending centrifugation, adding to a prepared suspension culture medium of the invention, carrying out beating-up into a single cell suspension, inoculating into a paved ultra-low absorption culture dish, and carrying out cell culture. The tumor stem cell suspension culture method has the following characteristics that: ball forming rates of various in vitro cultured tumor cells are high, multi-function factors are expressed, the OCT4 proportion and the NANOG proportion are increased, rich and comprehensive experimental materials are provided for tumor stem cell researches, and broad application prospects are provided in basic researches and clinical applications.

The invention discloses a method for culturing bladder cancer organs in vitro. The method comprises the following steps: manufacturing a gas-liquid interaction culture system, namely manufacturing a uniformly paved rat tail collagen supporting layer on the surface of a porous culture membrane in a Transwell upper chamber; re-suspending fresh in-vitro bladder cancer tissues by using a rat tail collagen solution, uniformly mixing, adding the obtained mixture onto the rat tail collagen supporting layer, and then putting the rat tail collagen supporting layer into an incubator at 37 DEG C to solidify so as to obtain a rat tail collagen layer containing the bladder cancer tissues; adding an organoid culture medium into a Transwell lower chamber, and enabling the liquid level of the culture medium to be lower than the rat tail collagen layer containing bladder cancer tissues; and carrying out passage and cryopreservation. The culture success rate of the bladder cancer organoid is remarkablyincreased, the bladder cancer organoid with reserved immune cells can be obtained through culture, operation is easy, the utilization rate of tumor tissue is high, and great significance and value areachieved for bladder cancer drug screening research.

The invention puts forward a universal cancer organoid in-vitro culture medium, which consists of a DMEM / F-12K (Dulbecco's Modified Eagle Medium / Nutrient Mixture F-12K) basic culture medium and an adding factor, wherein the adding factor consists of HEPES (2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid), L-Glutamine, EGF (epidermal growth factor), Noggin, FGF (fibroblast growth factor)-10, A83-01 and Y27632. According to the culture medium of the embodiment of the invention, the success rate of organoid passage culture can be greatly improved, in addition, the universal culture of cancer samples, including gastric carcinoma, rectal carcinoma, lung carcinoma and the like, can be realized, culture cost is greatly lowered, the long-term culture of the organoid can be realized, and abiobank is established.

The preparation process of cell-eliminating pig corium substrate includes sustained vibration of the corium substrate in 0.5 mol / L NaOH solution compounded with thrice distilled water, rinsing with thrice distilled water and balanced phosphate salt solution separately, freeze drying, packing and sterilizing. The cell-eliminating pig corium substrate is used as the in vitro corium culturing rack in composite skin preparing tissue engineering process, and the preparation process includes coating the corium substrate with acetic acid dissolved type-I ox collagen, regulating pH to 7.0-7.4, soaking subsequently in balanced phosphate salt solution and no serum culture medium, inoculating passage epidermal cell, and no serum culture inside a CO2 hatcher. The tissue engineering composite skin is used as human skin substitute.

The present invention relates to a method of isolating and culturing mesenchymal stem cells using umbilical cord blood that is most ideal for cell therapy. The method comprises adding an anti-coagulant to umbilical cord blood having a volume of more than 45 ml per unit, which is obtained within 24 hours after parturition; diluting the resulting mixture of the anti-coagulant and umbilical cord blood with an alphaMEM (alpha-minimum essential medium), followed by centrifugation to harvest monocytes; and subjecting the obtained monocytes into suspension culture in the alphaMEM containing Stem Cell Factor, GM-CSF (granulocyte-macrophage colony-stimulating factor), G-CSF (granulocyte colony-stimulating factor), IL-3 (interleukin-3) and IL-6 (interleukin-6).

The invention provides a bladder cancer organoid culture medium and a culture method. According to the culture method, two different organoid culture mediums are adopted for culture, and the composition and proportion of the two organoid culture mediums are disclosed. According to the method and the culture medium, the culture success rate, the passage frequency, the growth speed and the growth form of the bladder cancer organoid can be improved, and the problems that in the prior art, the culture success rate is low, the passage frequency is small, the organoid grows less, the cell form is poor, and growth is slow of the bladder cancer organoid are solved.

The invention discloses a method of cultivating Gastrodia elata and belongs to the technical field of planting of Gastrodia elata. The method comprises the steps of preparing Armillaria mellea, preparing mushroom sticks, inoculating, cultivating Gastrodia elata, and managing. By controlling the cultivation process of Armillaria mellea and modifying the method of cultivating Gastrodia elata, the yield of Gastrodia elata is increased; tubers of Gastrodia elata cultivated through the method are uniform in size, full and good in condition.

The disclosure relates to a method for culture of a cis-platinum drug-resistant lung cancer organoid. The method comprises the following steps: mixing cis-platinum drug-resistant lung cancer tissue cells with the medium and matrix gel to obtain a to-be-cultured substance, wherein the medium contains matrix metalloproteinase-10, and the concentration of the matrix metalloproteinase-10 is 50-200nM based on the medium; and carrying out culture amplification on the to-be-cultured substance to obtain the cis-platinum drug-resistant lung cancer organoid. According to the culture method for the cis-platinum drug-resistant lung cancer organoid provided by the invention, since the employed medium contains the matrix metalloproteinase-10 with specific concentration, and the matrix metalloproteinase-10 with the specific concentration can promote in-vitro growth of the cis-platinum drug-resistant lung cancer organoid, the success rate of the culture is high when the method provided by the disclosure is used for the culture of the cis-platinum drug-resistant lung cancer organoid.

The invention relates to a method for improving the success rate of umbilical cord blood mesenchymal stem cell culture by using placental mesenchymal stem cells. The method comprises the following steps: firstly, digesting placentallobules in a short time by using collagenase I to obtain a paste mixture consisting of extracellular matrix protein and tissue blocks and carrying out primary culture; and then co-culturing third-generation placental mesenchymal stem cells as moisturizing cells and separated primary umbilical cord blood mesenchymal stem cells under a low-oxygen condition to obtain a lot of umbilical cord blood mesenchymal stem cells. The method for improving the success rate of umbilical cord blood mesenchymal stem cell culture by using placental mesenchymal stem cells provided by the invention shortens the culture time of the placental mesenchymal stem cells, effectively solves the problem that the placental mesenchymal stem cells are low in purity and poor in multiplication capacity and the like, improves the success rate of umbilical cord blood mesenchymal stem cell culture and develops the utilization value of the umbilical cord blood mesenchymal stem cells. As fetal calf serums and pancreatin are not used, the safety of the cells on clinical application is enhanced.

The invention discloses a method for breeding, culturing and storing plant nematodes by using sweet potato calluses. The method is implemented through the following steps of: cleaning fresh sweet potatoes, performing surface sterilization, peeling, slicing, placing into a culture dish, and culturing in an incubator to obtain sweet potato calluses; inoculating sterilized plant nematodes onto the sweet potato calluses; placing into respective incubators of appropriate temperatures, and culturing and breeding in dark; and after culturing for a certain period of time, storing the plant nematodes and the sweet potato calluses at the temperature of 4-20 DEG C. Due to the adoption of the method, the aims of manually culturing and breeding a large quantity of plant nematodes in a simple way and storing plant nematode resources for a long time can be fulfilled.

The invention relates to a method for culturing liver cancer organoid. The method comprises the following steps: mixing liver cancer tissue cells with a culture medium and matrix gel to obtain a to-be-cultured substance, wherein the culture medium containing an insulin growth factor-2, and the concentration of the insulin growth factor 2 being 1-50ng / ml, preferably 5-10ng / ml based on the culture medium; and carrying out culture amplification on the to-be-cultured substance to obtain the liver cancer organoid. According to the culture method of the liver cancer organoid provided by the invention, the adopted culture medium contains the insulin growth factor-2 with the specific concentration, and the insulin growth factor-2 with the specific concentration can promotes in-vitro growth of theliver cancer organoid, so that the culture success rate is relatively high when the method provided by the invention is used for culturing the liver cancer organoid.

The invention discloses a seed treatment method for increasing the germination rate and improving the disease resistance of soybean seeds. The seed treatment method comprises the specific steps of seed pretreatment, preparation of seed soaking liquid, seed soaking, disease resistance treatment and sterilization treatment. The seed pretreatment comprises the following steps: putting mature soybean seeds into acidic soaking liquid with the temperature of 20 to 30 DEG C for 50 to 60 minutes, taking out the seeds, putting the seeds on sand paper, rinsing the seeds with clean water after wax on the outer skins of the seeds fall off, screening the seeds, and removing flat particles; the preparation of the seed soaking liquid comprises the following steps: adding the following components in weight proportion into a common aqueous liquid: 20 to 24g / L ammonium nitrate, 0.4 to 0.6g / L sodium phthalate, 0.3 to 0.5g / L sodium humate, 2 to 4mg / L nutrition adding liquid and 0.04 to 0.06mg / L gibberellin, and uniformly mixing for later use. The seed soaking liquid is enriched with nutritional elements, so that the soybean seeds can get through the period of dormancy within shorter time, the speed that the soybean seeds are germinated into seedlings is increased, irradiation of infrared achieves a sterilization effect; the seeds are soaked in potassium permanganate or formalin, the disease resistance of the seeds is enhanced, so that the culture success rate of the soybeans is increased, and high germination rate is realized.

The invention relates to a method for culturing a dedifferentiated or undifferentiated thyroid carcinoma organ. The method comprises the following steps: mixing dedifferentiated or undifferentiated thyroid carcinoma tissue cells with a thyroid carcinoma culture medium and matrix glue to obtain a to-be-cultured substance, wherein the thyroid carcinoma culture medium contains nicotinamide and BM-Cyclin, and based on the thyroid carcinoma culture medium, the concentration of nicotinamide is 5-20 mM, and the concentration of BM-Cyclin is 5-30 [mu]g / ml; and carrying out culture amplification on theto-be-cultured substance to obtain the dedifferentiated or undifferentiated thyroid carcinoma organ. In the method for culturing the dedifferentiated or undifferentiated thyroid carcinoma organ, theadopted thyroid carcinoma culture medium contains nicotinamide and BM-Cyclin with specific concentrations, so that the in-vitro growth of the dedifferentiated or undifferentiated thyroid carcinoma organ is promoted. Therefore, when the method provided by the invention is used for culturing the dedifferentiated or undifferentiated thyroid carcinoma organ, the culture success rate is relatively high.

The invention relates to a biomedical technology, in particular to a culture medium and culture method for ovarian cancer organoids. The culture medium comprises a basal culture medium DMEM / F12, B27, glutamine, HEPES, N-acetylcysteine, FGF2, A83-01, nicotinamide, forskolin, beta-estradiol, hydrocortisone, R-spondin-1, noggin, FGF10, EGF and Heregulin beta 1 and at least one selected from PFBS, PFOA and PFOS. According to the culture medium and the culture method, the establishment success rate of an ovarian cancer culture medium is improved, the amplification efficiency of the ovarian cancer organoids is improved, the proliferation efficiency after passage is stable, the drug screening process is accelerated, and the cost of culture materials is reduced.

The invention belongs to the technical field of plant cultivation and specifically relates to a medium for cultivating orchids and a preparation method thereof. The medium for cultivating orchids is prepared from the following raw materials: chestnut thorn, chestnut leaves or oak leaves, peanut shells and / or pine tree bark, gravel pebbles, rainwater and toshin. The orchid cultivated by the medium provided by the invention has the characteristics of excellent growth condition, dark green leaf color, strong root system, straight flower stalk, bright flower color, fragrance and long flowering phase; the replacing period of an orchid flowerpot is prolonged to 3-4 years; the success rate of orchid cultivation is increased, so that the living quality and grade of people can be promoted; and the cultivation cost of high-class flowers, such as orchids, can be lowered, so that new power is supplied for the development of the flower career.

The invention discloses a culture method of human colorectal cancer tissue organoid, which comprises the following steps: washing human colorectal cancer tissues and putting them into a transfer culture medium, cleaning and shearing; further cleaning, centrifuging, carrying out enzyme digestion, filtering, and collecting cell clusters; and mixing the cell clusters with hydrogel, adding DMEM culture medium to culture after solidification to obtain the human colorectal cancer tissue organoids. According to the invention, gentamicin, amphotericin B, primocin and other antibiotics are added into the transfer culture medium, the digestion solution and the culture medium, so that microbial contamination can be reduced, and the method for obtaining the human colorectal cancer organoids through in-vitro culture is simple, convenient, rapid, high in success rate and good in organoid growth state. The invention further discloses application of the human colorectal cancer organoid in drug testing, and the co-culture model of human colorectal cancer tissue organoids and tumor-associated fibroblasts can be used as a patient individualized drug testing platform.

The invention discloses a cultivation method of Gastrodia elata Bl. and belongs to the technical field of Gastrodia elata Bl. planting. The method includes the steps of preparation of a halimasch culture medium, preparation of bacteria sticks, inoculation, Gastrodia elata Bl. cultivation and management. By controlling the process of cultivating halimasch and improving a Gastrodia elata Bl. cultivation method, the aim to improve the yield of Gastrodia elata Bl. is achieved, and the Gastrodia elata Bl. prepared by the cultivation method is uniform in size, full, and good in appearance and quality.

The invention relates to a method for preparing an ovarian cancer organoid, which is characterized by comprising the following steps of: a, mixing feeder layer cells, temperature-sensitive hydrogel, ovarian cancer cells and a first culture medium, and culturing for 7-9 days to obtain a first culture; wherein the feeder layer cells are human endometrial stem cells without a proliferation function;and b, changing the liquid of the first culture by using the first culture medium, and culturing until the ovarian cancer organoid is formed. According to the method disclosed by the invention, the ovarian cancer organoid matched with ovarian cancer tissue biological information can be cultured by utilizing a small amount of ovarian cancer tissues or cells, and the culture success rate is relatively high.

The invention provides a culture medium, a culture method and a detection method of a thyroid cancer organoid, the culture medium comprises a basic culture medium, B-27, N-acetylcysteine, nicotinamide, R-spondin 1 recombinant protein and other components, the culture medium has high thyroid cancer organoid culture success rate, high passage frequency, and good passage stability after repeated cryopreservation and resuscitation; the thyroid cancer organoid obtained by adopting the culture medium and the culture method is high in reducibility; and the detection method is comprehensive and accurate.

The invention relates to the technical field of cell preservation and transportation, and particularly discloses a low-temperature preservation solution and a preservation method thereof. The low-temperature preservation solution is prepared from a DMEM high-glucose culture medium, Glutamax, HEPES and Penicilline-Streptomycin. The organoids transported by the cryopreservation solution at low temperature do not need to be resuscitated, and can be quickly cultured, is relatively low in cost and relatively high in culture success rate, and the activity of cells can be maintained for a long time. According to the invention, the technical problems of high hardware requirements, complex transportation procedures, high cost, low efficiency, low success rate of cryopreservation recovery, worry about cell growth conditions and the like are solved.

The invention provides a 3D culture system of primary tumor cells, and an application of the 3D culture system. The 3D culture system comprises lower layer gelatin, a middle layer cell substrate and an upper layer culture medium, wherein the middle layer cell substrate comprises improved agarose gel, a condition culture medium containing addition factors and tumor cells; the improved agarose gel comprises low melting-point agarose, polylysine and an extracellular matrix; the lower layer gelatin comprises agarose gel and a conditioned medium; and the upper layer culture medium is the conditionculture medium containing the addition factors. The 3D culture system provided by the invention can screen tumor cells and normal epithelial cells, besides, can realize preferential growth of the tumor cells, is high in culture success rate, can really reflect the properties of entire tumor cell groups, and can accurately represent the growth condition of in vivo tumors, and after the 3D culture system is used for medicine susceptibility detection, the obtained results are high in reliability, and clinical promotion is facilitated.

The invention relates to a portable anaerobion incubator. The portable anaerobion incubator comprises a box body, a single chip microcomputer, a control module, a display module, a power source circuit and a temperature and humidity sensor, a CO2 sensor, a heater and a moisturizer arranged in the box body; the control module, the display module, the power source circuit, the temperature and humidity sensor, the CO2 sensor, the heater and the moisturizer are all connected with the single chip microcomputer; the control module comprises a plurality of press buttons and a connection circuit enabling the multiple press buttons to be connected with the single chip microcomputer. The portable anaerobion incubator provided by the invention is scientific and reasonable in design, easy and convenient to operate, can conduct accurate and stable automatic control and adjustment on the temperature and humidity and the CO2 concentration in the box body, and guarantees that the conditions of the environment beneficial for bacteria to grow remain in the most proper range, the bacteria culture success rate is high, and the requirements for practical application can be well met.

The invention relates to a full-automatic multifunctional cell treatment system which comprises a biosafety cabinet and a workbench, and the workbench comprises an on-table part and an off-table part; wherein the on-table part of the workbench comprises a balancing unit, a heating unit, a quality control unit, a connection unit, a sample unit, a reagent unit, a consumable unit, a gun head unit, a waste unit, a liquid suction device, a first controller, a cap screwing device with recognition and clamping functions, and a second controller; the off-table part of the workbench comprises a centrifugal machine and a power supply control module. The biosafety cabinet is arranged outside the workbench, and a switch window is arranged on the biosafety cabinet.

The invention discloses a leaf cutting propagation method of succulent plants of xPachyveria pachytoides, and belongs to the technical field of a planting propagation method. The method provided by the invention is characterized in that a leaf of a xPachyveria pachytoides mother plant subjected to shade still standing is pulled and taken, and is then subjected to air drying for drying the broken edge; then, the back side of the obtained leaf is in contact with culture soil prepared in advance, and is then subjected to indoor scattered light culture; when roots being 5mm grow from the leaf, planting soil is used for covering the root parts; after the number of the leaves exceeds eight, and the plant reaches the height of 2cm, the foliage fertilization is performed; after the plant continuously grows by at least 1cm, the transplanting is performed, wherein the culture soil is prepared by uniformly mixing sand, coal slag and peat soil which are dried in the air for 2 to 3 days and have the same weight. The method provided by the invention has the characteristics that the operation is simple; the propagation period is short; the survival rate is high, and the like. The method is very suitable for large scale culture.

The invention discloses a method for cultivating gastrodia tuber by clinker wood. The method comprises the following steps: taking the raw materials of 90-110 parts of raw material saw dust, 50-70 parts of wheat bran, 15-25 parts of corn flour, 1-3 parts of brown sugar and 1-3 parts of plaster powder; evenly mixing the raw materials to obtain a mixed filler for backup; segmenting the wood into 15cm-small-wood sections; binding the 15cm-small-wood sections and then loading into a strain bag, filling up gaps in the strain bag with the obtained mixed filler, and then fastening the opening of the bag; placing the bound strain bag into a sterilizing pot for sterilization; heating to 100 DEG C under the condition of normal-pressure sterilization and then keeping for 8-10 hours; after stopping fire, keeping for 2 hours, and then opening an oven door for heat dissipation; cooling the temperature of the bag to 70 DEG C, getting the bag out of an oven, and then transferring the bag into a cooling room; and finally cooling, inoculating, culturing the wood indoors, and cultivating outdoors. The method of the invention carries out high-temperature disinfection treatment on the wood, thus achieving less mixed wood pollution, short wood cultivation time, high culture success ratio of the wood and hypha and saved resources.

The invention discloses a primary culture method of the embryonic cell tissue pieces of a Chinese softshell turtle. The primary culture method comprises the following steps of: (1) collecting the fertile eggs of the Chinese softshell turtle, and disinfecting; (2) breaking the eggshells of the fertile eggs, separating embryos from yolk, cleaning the separated embryonic tissues with a 1*PBS (Phosphate Buffer Solution), and then soaking for sterilization; (3) cleaning the embryonic tissues for 4-5 times in an M199 culture medium, then transferring into a complete culture medium, cutting the embryonic tissues into pieces to obtain tissue pieces, and then cleaning the tissue pieces once by using the complete culture medium; (4) uniformly dispersing the tissue pieces to the inner wall of a cell bottle by using a sterile disposable inoculating needle so that the tissue pieces grow for 3-4 hours in a way of being attached to the wall, then adding the complete culture medium to submerge the tissue pieces, and culturing at the constant temperature of 30-31 DEG C for more than 3 days. The primary culture method disclosed by the invention has the advantages of easy operation, convenient culture, no need of specific cell separation liquid and high culture success rate.

The invention relates to a culture medium of amniotic fluid cells, which belongs to the field of biochemistry. The culture medium comprises the following components: basic medium, transferrin, sodium selenite, insulin, triiodothyronine, glucagons, basic fibroblast growth factor, hydrocortisone, testosterone, estradiol, progesterone and added antioxidant, wherein the antioxidant is a mixture of vitamin E and L-ascorbic acid; and the culture medium can be used in amniotic fluid cell culture after 4 to 15 volume percent of fetal calf serum is added in the culture medium, and a great amount of amniotic fluid cells which are at a division stage and meet the requirements of clinical diagnosis and scientific research can be obtained just within 7 to 8 days. The culture medium remarkably improves the culture effects of amniotic fluid cells and obviously increases cloning efficiency and cloning growth rate, reduces culture time, increases the success rate of cell culture, and fully plays the synergetic effects of vitamin E and L-ascorbic acid of different antioxidants.