75 results about "Explant culture" patented technology

The invention discloses a stem cell tissue culture seedling raising technique. The technique comprises the steps of 1, taking a fresh plant, conducting sterilization on the surface and cutting the plant; 2, conducting tissue differentiation and reserving a cambium; 3, culturing the cambium on an isolation medium. According to the technique, existing methods are improved based on a traditional tissue culture technique, plant stem cells are taken as a target, induction, separation and explant culture are conducted, and a corresponding stem cell culture system is established. By the adoption of the plant stem cell culture technique, the plant culture technique is enriched, and a novel research direction and opportunity is provided for commercialized production of natural products and development of the plant biotechnology.

The present invention relates to an efficient method for isolating and culturing mesenchymal stem cells from umbilical cords, i.e. a staged suspension explant culture method, comprising the following steps: (1) digesting an umbilical cord in vitro by using a tissue digestive enzyme; (2) culturing the digested cells with a mesenchymal stem cell culture medium for culturing to obtain mesenchymal stem cells; (3) culturing the tissue being not fully digested after digestion by a mesenchymal stem cell culture medium to obtain the mesenchymal stem cells. According to the present invention, the mesenchymal stem cells obtained by the method are in line with mesenchymal stem cell characteristics in surface markers, growth characteristics, multi-directional induced differentiation and other aspects. The method is simple, a lot of mesenchymal stem cells could be obtained at the same time, and the method provides a new way for obtaining mesenchymal stem cells.

The invention relates to a stevia rebaudiana tissue culture propagation method and a culture medium of the method, and particularly relates to a tissue culture rapid propagation method for directly forming a bud by using a stevia rebaudiana leaf as an explant. The stevia rebaudiana tissue culture method comprises the following steps: (1), collecting the explant; (2), disinfecting and inoculating; (3), performing thickening induction-culturing; (4), budding and seedling; (5), transferring and grafting, and rooting; and (6), hardening and transplanting. According to the stevia rebaudiana tissue culture method provided by the invention, materials are easily available, a plurality of seed sources can be rapidly propagated, the explant tissue culture has no culture phase of callus cell proliferation and differentiation, thus the culture time is shortened, extremely high reproduction rate is achieved; the stevia rebaudiana tissue culture propagation method is easy to operate and is capable of realizing large-scale production; and the operation of researches such as genetic transformation is facilitated in a culture process. The culture medium provided by the invention ensures that the leaf is capable of reaching the expected culture target in each phase.

A method is disclosed for producing a transgenic cotton plant comprising the steps of (a) obtaining cottonfibrous root explants, (b) culturing the fibrous root explants to induce callus formation, (c) exposing root callus to a culture of Agrobacterium tumefaciens that harbors a vector comprising an exogenous gene and a selectable marker, the Agrobacterium being capable of effecting the stable transfer of the exogenous gene and selection agent resistance gene to the genome of the cells of the explant, (d) culturing the callus in the presence of the selection agent to which the selection agent resistance gene confers resistance so as to select for transformed cells, (e) inducing somatic embryo formation in the selected callus culture, and (f) regenerating the induced somatic embryos into whole transgenic cotton plants.

The invention relates to a method for high-salt screening of a saline-alkali resistant rosa chinensis plant. In particular, rosa chinensis which is planted in Tibet plateau is obtained by the method for high-salt screening of saline-alkali resistant rosa chinensis. The method comprises the steps of inducing obtained rosa chinensis explant culture medium suitable for rosa chinensis explants out of a sprout texture and then irrigating and screening by a high-salt culture medium to obtain the saline-alkali resistant rosa chinensis; cutting a fibroid branch with thick vascular bundle into 10cm stem sections, sterilizing the culture mediums by 0.5% potassium permanganate according to the ratio of vermiculite to perlite to nutrient soil being 10:2:1, and then spraying the medium by 0.3% rooting liquid, wherein high-salt screening is diluting and irrigating by sodium chloride liquid culture mediums B with different concentrations, and selecting the plant which can normally grow. By adopting the method, the growth boundaries of fresh flowers are broken through; five saline-alkali resistant rosa chinensises can be obtained from genetic modification and individual gene screening, and a blank no fresh flower is available in golmud is filled.

The invention provides a method for tissue culture and rapid propagation of chimera hosta plants, which includes disinfection, primary culture, subculture and rooting culture, and utilizes young buds of chimera hosta as explants for tissue culture and rapid propagation. The method has a high reproduction coefficient, is suitable for commercial production, and fills a gap in market demand for chimera hostas. The present invention uses the young shoots of the chimera hosta as explants, and studies the plant regeneration and rapid propagation technology of the hosta flower. The whole industrialization process of plant regeneration, multiplication, rooting and seedling formation is completed by the invention, and the high proliferation coefficient of the culture medium can meet the large demand of commercial production.

The invention is directed to a chemically defined animal cell culture media, and methods for preparing such a medium, wherein the media are suitable for culturing epidermal cells, preferably human epidermal cells, including cells of the hair follicle. The invention further provides for methods of culturing epidermal cells, hair follicles, and skin explants in the media as well as uses of the cell cultures and explant cultures in screening assays.

The invention discloses a method for quickly propagating plukenetia volubilis linneo. The method includes steps of disinfecting the surfaces of explants which are current-year non-lignified branches of the plukenetia volubilis linneo; cultivating and inducing callus tissues of the explants; inducing differentiation and elongation of buds by the aid of the callus tissues; strengthening and rooting seedlings; exercising and transplanting the seedlings and the like to obtain regenerated plants. The method has the advantages that integral procedures can be carried out under controllable conditions without seasonal constraints, the propagation cycle can be shortened to a great extent, the propagation multiples can be increased, the method has important practical significance on meeting requirements on high-quality seedlings of the plukenetia volubilis linneo and accelerating popularization of improved varieties of the plukenetia volubilis linneo, and an efficient and feasible technical means can be provided for future research on genetic modification and the like on the plukenetia volubilis linneo.

The present invention features a human skin explant culture system and uses thereof.

The invention relates to preparation and storage of mesenchymal stem cells for clinical treatment. The preparation method comprises the following steps: storing a physiological saline umbilical cord containing 0.2% of genramicin and 0.54% of anticoagulant; shearing the umbilical cord to 2-3cm; centrifuging, suspending, and centrifuging; directly adhering and carrying out explant-culture; adding an alpha-minimum essential medium (MEM) containing 15% of fetal calf serum and 0.2% of genramicin, and culturing; after cell fusion, digesting the cells with digestive juice containing 0.05% of pancreatin / 0.2mM ethylene diamine tetraacetic acid (EDTA); carrying out passage in a ratio of 1:3 to 4-5 generations, so as to obtain a large amount of mesenchymal stem cells; and putting a frozen storing liquid (the ratio of medium to fetal calf serum to dimethylsulfoxide is 5:4:1) in the mesenchymal stem cells, placing in a refrigerator of minus 70 DEG C overnight and then storing in liquid nitrogen, wherein the freezing temperature of the liquid nitrogen is minus 196 DEG C.

The invention discloses a propagation method of oriental lily. The propagation method comprises the following steps: taking thrums of the oriental lily as explants, sterilizing the thrums, inducing calluses, inducing adventive buds, proliferating the adventive buds, inducing the adventive buds to root, hardening and seedling tissue culture seedlings, transplanting the tissue culture seedlings and the like until the young plants are formed. The method is significant to lower the production cost of the oriental lily and improve the quality of lily. The method relates to selection of oriental lily explants and the sterilization method, selection of plant hormones, and optimization of induction conditions of the calluses, induction of the adventive buds and proliferation conditions, rooting culture conditions, hardening-seedling, transplanting and other conditions. The method has the characteristics of high simplicity and good feasibility in explant sterilization method, wide explant culture conditions, low culture difficulty, and high survival rate of tissue culture seedlings, and serves as a simple, efficient and quick propagation method for the oriental lily.

The invention discloses an inducing method of dioscorea alata transgenosis hairy roots, and belongs to the technical field of plant culture and transgenosis. The method includes the steps: 1) explantsterilization; 2) explant culture; 3) infection liquid preparation; 4) hairy root induction and reproduction; 5) hairy root reproduction. According to the method, dioscorea alata stems with leaf axilla serve as explants, an induction culture medium, a differential culture medium and a rooting culture medium are optimized, an integrated plant is acquired through induction, differential and a rooting culture, stems of dioscorea alata aseptic seedlings are infected by agrobacterium rhizogenes K599 (with recombinant plasmid pRI101-AN-gfp) living bodies to obtain the transgenosis hairy roots, PCR (polymerase chain reaction) and fluorescence microscope observation identify whether the hairy roots are transgenosis hairy roots or not, an inducing system of the dioscorea alata transgenosis hairy roots is built, and dioscorea alata is rapidly and effectively bred.

The invention relates to a method for cultivating thornless Lycium ruthenicum Murr.. The cultivation method is characterized by comprising the following steps: by using the stem or mature seed of thorny Lycium ruthenicum Murr. as the original donor material, performing culture by using the stem or leaf as an explant to obtain a sterile tissue culture seedling; inoculating the obtained tissue culture seedling into an improved 1 / 4 or 1 / 2 MS solid culture medium, and performing strong seedling culture under the sunlight; after the sound seedling culture, performing transplanting and acclimatization; putting the newly transplanted tissue culture seedling under scattered light, and periodically pouring distilled water to keep the water content of the culture medium at the field moisture capacity of 100%; performing water control treatment on the healthy potted tissue culture seedling which survives after the transplanting and acclimatization, and periodically pouring a 1 / 8 improved MS nutrient solution or 1 / 4 improved MS nutrient solution; and putting the potted seedling under the sunlight, and keeping the ambient temperature at 18-28 DEG C. The thorny Lycium ruthenicum Murr. is utilized as the starting material to successfully cultivate abundant thornless Lycium ruthenicum Murr. plants, and the thornless plants can grow and develop very well, thereby laying the foundation for the cultivation of new species of thornless Lycium ruthenicum Murr..

The invention belongs to the field of agricultural high technologies and specifically relates to a hemsleya amabilis tissue culture and lateral propagation method. The method comprises the following steps of initial medium preparation, explants sterilization and treatment, inoculation and culture, subculture medium preparation and inoculation, rooting culture, seedling hardening and transplantation. The method disclosed by the invention overcomes loss of effective ingredients cultured under a specific environment of a laboratory in the prior art and achieves artificial cultivation of wild varieties. Detection shows that dihydrocucurbitacin, cucurbitacin and hemsleya amabilis saponins all can be detected from the hemsleya amabilis propagated by the method disclosed by the invention, and contents of the dihydrocucurbitacin, the cucurbitacin and the hemsleya amabilis saponins are equivalent to that of the dihydrocucurbitacin, the cucurbitacin and the hemsleya amabilis saponins in wild hemsleya amabilis. The method disclosed by the invention improves a qualified seedling percentage of hemsleya amabilis seedlings, shortens a cultivation period, keeps effectiveness of the wild hemsleya amabilis and has an application prospect in the aspects of preserving, protecting, developing and utilizing wild hemsleya amabilis resources and the like.

The present invention provides methods for individualizing chemotherapy for cancer treatment, and particularly for evaluating a patient's responsiveness to one or more epidermal growth factor receptor (EGFR) inhibitors prior to treatment with such agents. Particularly, the invention provides an in vitro chemoresponse assay for predicting a patient's response to a monoclonal EGFR antibody, such as cetuximab. The method generally comprises culturing malignant cells from a patient's specimen (e.g., biopsy specimen), contacting the cultured cells with a monoclonal EGFR antibody that is a candidate treatment for the patient, and evaluating the cultured cells for a response to the drug. In certain embodiments, monolayer(s) of malignant cells are cultured from explants prepared by mincing tumor tissue, and the cells of the monolayer are suspended and plated for chemosenstivity testing. The in vitro response to the drug as determined by the method of the invention is correlative with the patient's in vivo response upon receiving the monoclonal EGFR antibody during chemotherapeutic treatment (e.g., in combination with other standardized or individualized chemotherapeutic regimen).

The invention relates to a method for rapidly propagating sugarcane tissue culture seedlings with spermine. The method comprises the following steps: using sugarcane immature leaf slices as explants, vaccinating the explants to an improved MS solid medium of MS+Spm.1.5 mg . L<-1>-3.0mg . L<-1>, regulating the pH to be 5.8-6.0, and performing continuous culture for 10 d-25 d to induce and obtain sugarcane callus tissue agglomerates; retransferring the agglomerates to the same fresh culture medium for subsequent culturing for 15 d, and then separating and cutting obtained callus tissues into small blocks to be revaccinated to the same fresh culture medium; performing subsequent culture for 20d -30d, transferring regeneration seedlings to the same fresh culture medium to be cultured for 20d so as to obtain test-tube seedlings of which the stem length is 5-6cm and the root seedlings are complete. According to the invention, only one culture medium is used from the starting of explant culture to the generation of the complete regenerated plantlets, so that the production process for the sugarcane tissue culture seedlings is obviously simplified, besides, the toxic action of 2,4-D induction callus tissues is removed, and the cost of the used spermine is low, so that the method is more favorable for popularization.

The invention discloses a method for lily virus-free cultivation through using moroxydine drug, belonging to the fields of plant culture rapid propagation technique and biotechnology. The method comprises the steps of detecting lily seed balls virus, preparing culture medium, standby, sterilizing and vaccinating explants, culturing the explants and detecting tissue culture seedling virus, etc so as to obtain a large number of virus-free tissue culture seedlings quickly. The best induction culture medium formula that lilies are directly divided into sprouts and root-free tissue culture seedlings is obtained through an orthogonal method; the optimal virus-free concentration is screened through preparing a series of moroxydine antiviral drugs with concentration gradient; the virus-free efficiency can reach 91%; the induction culture period is shortened; the method has the characteristics of high virus-free efficiency and high enhancement factor, etc; and the method offers technological base to culture lily virus-free plants and popularize virus-free cultivation research.

The invention discloses a method for inducing hairy roots of sophora tonkinensis roots. The method comprises the following steps: (1) disinfecting sophora tonkinensis seeds, and then inoculating to an MS seed culture medium for germinating; (2) cutting the germinated seeds into chips of 1-2cm according to cotyledons, hypocotyls, leaves and stems, scratching with a scalpel, and inoculating to an MS explant culture medium for pre-culturing; (3) soaking the pre-cultured material into rooting agrobacterium liquid for bacterial infection; (4) inoculating the material subjected to bacterial infection to an MS co-culture medium for co-culturing; and (5) inoculating the co-cultured material to a bacteriostatic culture medium for bacteriostatic culture and sterilization. By adopting the method for inducing the hairy roots, the obtained hairy roots have the characteristics of high growth speed, good genetic stability, strong synthetic capability and capability of stably releasing part of metabolites to the culture medium; and the method is simple and convenient in operation, fast and high in induction rate, and can be used for producing industrial replaceable resources of sophora tonkinensis roots in large scale.

The present invention provides methods for individualizing chemotherapy for cancer treatment, and particularly for evaluating a patient's responsiveness to one or more epidermal growth factor receptor (EGFR) inhibitors prior to treatment with such agents. Particularly, the invention provides an in vitro chemoresponse assay for predicting a patient's response to an EGFR inhibitor, such as an EGFR tyrosine kinase inhibitor or a molecule targeting the extracellular domain of EGFR. The method generally comprises culturing malignant cells from a patient's specimen (e.g., biopsy specimen), contacting the cultured cells with an EGFR inhibitor that is a candidate treatment for the patient, and evaluating the cultured cells for a response to the drug. In certain embodiments, monolayer(s) of malignant cells are cultured from explants prepared by mincing tumor tissue, and the cells of the monolayer are suspended and plated for chemosenstivity testing. The in vitro response to the drug as determined by the method of the invention is correlative with the patient's in vivo response upon receiving the EGFR inhibitor during chemotherapeutic treatment (e.g., in combination with other standardized or individualized chemotherapeutic regimen).

A process for quickly culturing transgenic plant of potato includes such steps as agroinfecting the potato explant cultured in test tube, culturing to introduce the target gene to potato receptor, direct culturing in solid differential culture medium to induce resistant buds, culturing in selective rooting culture medium to induco normal root, and verifying by PCR, PCR-southern, or Northern hybridizing method.

The invention relates to a method for obtaining hairy roots of broccoli. The method comprises the step (1) of clipping explants of aseptic seedlings of rapid propagation of the same batch, putting the explants on sterile filter paper, using a sterile needle head to prick 2-8 wounds on the surfaces of the explants, inoculating the explants in an MS culture medium and performing preincubate in the dark to obtain the oreincubate sterile seedling explants; step (2) of using agrobacterium rhizogenes to infect the sterile seedling explants to be cultured, using the sterile filter paper to soak up residual bacterial liquid on the surfaces, inoculating the explants to an MS culture medium adding acetosyringone to culture the explants together in the dark to obtain receptor explants cultured together; and step (3) of transferring the receptor explants cultured together to an MS culture medium adding Carbencillin Disodium to be cultured in the dark, transferring the receptor explants once every 5 days and obtaining the aseptic hairy roots after the explants are transferred from 2 to 5 times. The method is simple and efficient and suitable for industrialized production.

A device is provided for securing a tissue sample from biological material. The tissue sample is housed in bottom and top platens that are configured to promote fluid communication between the tissue sample and the exterior environment to permit transport or cryogenic fluid to contact the sample. Additionally, diskette assemblies may be provided within the platens that permit sub-samples to be separated without directly handling the tissue sample. The diskette assemblies may also be configured to promote fluid communication with the sub-sample housed therein.

The invention relates to a method for regenerating adventitious buds by inducing callus tissue through cotyledons of a herbaceous peony. The method comprises the steps of: initiating cultivation by using herbaceous peony seed embryos, disinfecting seed embryos, cutting the embryos, inoculating the cut seed embryos to a basal culture medium of an MS solid medium, performing cultivation for 15 daysto obtain embryo seedlings, cultivating strong seedlings for 20-30 days to obtain tissue culture seedlings which grow robustly, and performing induced cultivation of the callus tissue for 45-60 days to obtain a callus tissue block which is plump, fresh and green and has a compact structure; and transplanting the callus tissue block into an adventitious bud differentiating culture medium to obtainthe adventurous buds which grow robustly. According to the method for regenerating the adventitious buds by inducing the callus tissue through the cotyledons of the herbaceous peony, the raw materialsare convenient to collect, the seed embryos are adopted as the tissue culture seedlings of explant culture, no limitation is caused by seasons, and the raw materials can be collected all the year; the contamination rate of first-generation embryo culture is far lower than that of other explants, and the induction rate of the callus tissue and the differentiating rate of the adventitious buds areimproved greatly; in addition, through the addition of an anti-browning agent in the phase of callus tissue induction and adventitious bud differentiation, browning can be inhibited effectively, and the problem of high contamination rate during induction of the callus tissue by the explants of the herbaceous peony can be solved.

The invention discloses a method for culturing and producing micro-bulbodium of crocus sativus L. through a superficial layer. The method comprises steps: (1), sterilization of a culture container; (2), acquisition of an explant, and sterilization treatment; (3), preparation of the explant cultured on the superficial layer; (4), superficial layer cultivation, wherein the superficial layer cultivation stage comprises two processes of (A) cluster bud induction and (B) micro-bulbodium formation. A liquid culture mode is adopted for superficial layer cultivation, a culture body can more easily absorb nutritional ingredients in a culture medium, accordingly, seedlings can grow fast, and the production time of micro-bulbodium is shortened; meanwhile, the cultivation operation of the superficial layer is simple, the cost is low, and large-scale and industrial production is easier to realize.

The invention relates to a method using Na2S2O3 (sodium thiosulfate) to of preventing browning of a tobacco explant by using Na2S2O3 (sodium thiosulfate). The method comprises the following steps of disinfection of plant seeds, culturing of the plant seeds, culturing of the plant explant, anti-browning culturing of the plant explant, and subculture type anti-browning culturing of the plant explant, so that the browning rate of the plant explant is 18.7 to 23.3%. The method has the advantages that the browning of the explant is effectively relieved, and the whole differentiation state of the explant can be improved; the Na2S2O3 and natural anti-oxidizing components are added into a culture medium; the convenience in use is realized, the cost is low, the effect is good , and the like.

The invention relates to the technical field of plant culture and discloses a planting method of pearl plums. The planting method of the pearl plums disclosed by the invention has the beneficial effects that a tissue culture method is selected for raising seedlings of the pearl plums, so that the seedling raising time can be shortened; in seedling raising, a disinfectant prepared from plant extracts is used for replacing common chemical disinfectants, and meanwhile in the process of explant culture and rooting-seedling culture, a culture medium I, a culture medium II, a planting culture mediumand leaf fertilizer are prepared according to the growth characteristics of explants, rooting seedlings and young seedlings, so that the germination rate and the survival rate of the explants and therooting seedlings can be effectively increased and the quality of the young seedlings can be effectively improved; the seedling raising time can be shortened; organic fertilizers, organic water fertilizer and insect-repellent water are also prepared according to the characteristics of young tissue-culture seedlings to plant young pearl-plum seedlings, so that the growth of the pearl-plum trees can be promoted, and meanwhile no pollution can be caused to the environment.

The invention relates to the technical field of plant cultivation study accessory devices, in particular to a vegetative propagation explant culture device for rare plants. The work modes of the vegetative propagation explant culture device are diversified, the cultivation temperature and oxygen content in a containing cavity can be adjusted conveniently, the use limitation of the vegetative propagation explant culture device is reduced, the cultivation effect is good, and high practicality is achieved. All waste culture solution in the containing cavity can be drained conveniently, it can beavoided that part of the waste solution remains to influence the cultivation effect in the containing cavity, and therefore the use reliability of the vegetative propagation explant culture device isimproved. A storage rack is included, the containing cavity is formed in the storage rack, and a taking opening is formed in the top end of the storage rack. An aerating device, an electric heating bar, a refrigerator, a compressor, an evaporator, a condenser and a capillary tube are further included, and a ventilating pipe is arranged at the output end of the aerating device. A bottom plate, a hydraulic device, a first fixed frame, a second fixed frame, a first fixed shaft, a second fixed shaft, a servo motor, two movable plates and a chain are further included, and two fixed plates are arranged in the two fixed tanks respectively.

The invention discloses a cutting propagation technique for hydrilla verticillata, comprising the following steps: selecting and processing explants; preparing hormone medicine, processing explants and culturing explants in fields. When the explants not processed by hormones is cultured for one week, the rooting rate is 67% and the survival rate is 46%; the rooting rate of the explants processed by the hormones is 95% and the survival rate is 90%; and the survival rate is greatly improved. The hydrilla verticillata is propagated in a large scale by the method; the method for improving propagation survival rate of hydrilla verticillata can solve the problem of low propagation speed of hydrilla verticillata under natural conditions and meet the requirement on hydrilla verticillata seedlingswhen recovering aquatic vegetation and water eutrophication management.