73 results about "Developmental biology" patented technology

The present invention relates to compositions comprising a decellularized tissue. The present invention also provides an engineered three dimensional lung tissue exhibiting characteristics of a natural lung tissue. The engineered tissue is useful for the study of lung developmental biology and pathology as well as drug discovery.

A method for preparing paraffin section of seawater roe includes selection of fixing agent, confirmation of dewatering process, confirmation of transparent process, confirmation of wax penetration process, improvement of dyeing mode. In the method, egg capsule stripping process being difficultly operated is omitted.

The invention put forwards an excised cultivation technology of procambarus clakii eggs according to developmental biology principle. The key points of the technology are the separation between parent prawns and eggs of berried prawns, the classification of the eggs, and respectively artificial hatching of the eggs. The steps of the invention are as follows: firstly, the berried prawns are sterilized; secondly, the parent prawns and the eggs are separated; thirdly, the eggs in different development stages are classified; fourthly, nidation of the eggs; fifthly, the hatching of the eggs. The invention has a high egg hatching rate, uniform sized young hatching prawns and a high commodity value. The cultivation technology can effectively prevent the young prawns from eating each other; therefore the survival rate of the young prawns is improved. Because the berried prawns do not need to be reared, cultivation cost is reduced and the transportation of prawn eggs is more convenient.

The invention provides a method for reliably storing pear pollen germplasm for long term, belonging to the field of plant developmental biology. The method is characterized by putting the pear pollen into an allochroic silicagel box after drying the pear pollen, packing and sealing the box with tinfoil, freezing the pollen in liquid nitrogen and storing the pollen at ultralow temperature, placing the stored pollen for 24 hours at minus 73 DEG C, minus 20 DEG C and 4 DEG C respectively in sequence to gradually revive the pollen when the pollen needs to be used in the coming year and finally restoring the pollen to room temperature in dark environment for later use. The method is secure and reliable, can effectively maintain the vigor of the pear pollen, is simple and easy to operate, saves space, can be used for reference to preserve other germplasm resources and has wide application value.

The invention provides a flexible high-flux cell electric fusion microelectrode array chip device, which is composed of an array chip and a fusion pool. The array chip takes a flexible transparent polyimide film as a substrate, the lower surface forms a cross comb-shaped array multimicroelectrode through etching, the electrode group is composed of two comb-shaped microelectrode array electrodes which are mutually crossed but not contacted or connected in electrical structure, and a microchannel between both internal electrodes of the electrode group serves as a service passage; the array chipis inversely buckled on the fusion pool, and the cross comb-shaped array multimicroelectrode on the array chip corresponds to the cell electric fusion pool and falls in the cell electric fusion pool.The invention has the advantages that the use is convenient, the processing method is simple, the cost is extremely inexpensive, and the chip device can not cause damages to operators or cells, thereby being suitable for being widely applied in the fields of genetics, animal and plant distant hybridization breeding, developmental biology, medicine screen, mono-clonal antibody preparing and mammalian clone.

The invention relates to chicken EPGCs single cell cloning method. Its feature is that using the fourth generation chicken EPGCs to prepare single cell cloning; culturing and passage to gain heredity uniform chicken EPGCs cell. The invention has simple and practical method, no need special apparatus demand, uses regular cell culture to finish single cell cloning, processes morphology and staining identification for the gained cell to improve that it has its primary characteristic. The produced cell can be directly used in developmental biology study, genome imprinting study fields etc. Because of the cell has little heredity changeability, it can be used to study its different affecting factors.

The invention relates to a method for conveniently and fast observing stem tip flower bud differentiation of tobacco plant, belonging to the technical field of developmental biology of plants. The method comprises the following steps: a. taking a sample with a blade at the position which is 5 centimetres downward from the stem tip; b. after sampling, immediately placing the stem tip in an ordinary injector for medical use, which has a volume of more than 15mL, pushing a pushing rod of the injector to the bottom, using a thumb to block the front part of a needle tube and forcefully pulling up the pushing rod to the top of the injector; retaining the position for 1-3min and repeating the process for 3-4 times; c. fixing the stem tip sample in a stationery liquid for 24h for testing with components having the following proportioning: 5.0mL of formaldehyde with a concentration of 38%, 5.0mL of acetic acid, 90.0mL of alcohol with a concentration of 75%; d. using a stereomicroscope to observe and take photos of the sample. The method has the advantages of low cost, simple and convenient operation, fast observation and acquisition of digitalized pictures with clear images, thus being of great guiding significance for experimental studies.

The invention discloses a method for differentiating induced pluripotent stem (iPS) cells into cartilage cells. The method is characterized in that the iPS cells are subjected to induction culture by use of a conditioned medium in a cell micell mode, wherein in the medium, the high-glucose DMEM (Dulbecco Modified Eagle Medium) contains 5-10% of fetal bovine serum, 6-6.5mug/ml insulin, 10-20ng/ml transforming growth factor beta1, 95-110mumol/ml dexamethasone, 37-38mg/ml ascorbic acid, 0.8-1.2mumol/ml sodium pyruvate and 6-6.5mug/ml transferrin. The method disclosed by the invention is advanced in technology, simple in operation and strong in repeatability, and provides a chance for implementing the cell replacement therapy and finally realizing regeneration and replacement of the tissue and organ; and a cell platform for large-scale production is expected, and then a brand new research prospect is developed for the fields of development biology, medicine and genetics.

The invention relates to a method for separating and purifying a plant tapetum tissue, and belongs to the fields of developmental biology and cell biology. The method comprises the steps of collection of anthers, preparation of a cleaning liquid, preparation of an enzymolysis liquid, pretreatment of the anthers, removal of anther microspores, and separation and collection of tapetum cells. Suitable enzyme species and concentration are used for enzymolysis separation of a tapetum tissue, and then the tapetum is collected through filtration, centrifugation and purification. The method has the advantages of simple steps, convenient operation, shorter consumed time and the like, the separation efficiency is good, and the obtained tapetum tissue can be used for subsequent experiments.

The invention relates to a cell electrofusion chip device based on a surface microelectrode array and a deformation membrane structure. The cell electrofusion chip device is composed of a surface microelectrode array chip and a top-layer deformation membrane structure, wherein the surface microelectrode array chip is divided into a quartz base layer, a microelectrode array layer and a polymer microchannel layer; the microelectrode array is in a sandwich structure and is attached to the polymer microchannel layer, thereby avoiding the cell blockage problem caused by the traditional tooth-shaped projection electrode structure and simultaneously guaranteeing the integration level and better fusion efficiency of the microelectrode; the deformation membrane structure is composed of a flexible deformation membrane and an air pressure adjusting device; the flexible deformation membrane covers the microchannel; and the flexible deformation membrane deforms under the action of air pressure applied by the air pressure adjusting device, protrudes downwards and enters the microchannel to form a partition wall, thereby dividing the microchannel into two microchannels and realizing independent feeding and pairing of different cells. The device can be widely applied in the fields of genetics, distant hybridization breeding of animals and plants, developmental biology, drug screening, monoclonal antibody preparation, mammal cloning and the like.

The invention provides an expression promoter of rice stamens and lodicules. The expression promoter comprises a nucleotide sequence as shown in the sequence table SEQ ID No 1 or 2 or a derivative thereof. The invention further provides a recombinant expression vector, an expression cassette and host bacteria, which are constructed by using the promoter. The expression of an exogenous gene in the rice stamens and lodicules can be driven by linking the expression promoter of rice stamens and lodicules to a gene to be expressed and transferring the gene into rice plants by a vector. The promoter specifically expressed in stamens and lodicules may be a useful tool in the research of rice fertilization. With the development of molecular biology and developmental biology, the demand for such a specific expression promoter becomes strong increasingly, and the expression promoter of rice stamens and lodicules just meets the demand. The invention has a huge potential commercial value in the cultivation of male-sterile plants, breeding and other aspects, especially.

The invention provides a Cyprinus carpio var. jian cerebral line and application thereof. Through primary isolation on Cyprinus carpio var. jian cerebral tissues with pancreatin of an appropriate concentration and reasonable digestion conditions, Cyprinus carpio var. jian cerebral cells can be obtained, and through subculture, the Cyprinus carpio var. jian cerebral line CCB-J is established for the first time; the Cyprinus carpio var. jian cerebral line is mainly represented as epithelioid cells and can be passaged for more than 60 generations, thereby be high in research and application the fields such as virology, toxicology, physiology, molecular genetics and biology of evolution. The Cyprinus carpio var. jian cerebral line can be applied to virus identification and virus strain isolation, to preparation of Cyprinus carpio var. jian virus preventing or killing vaccines, as biological models of drug screening, preparation and evaluation and the like.

The invention discloses a method for separating and primary culture of zebrafish intestinal mucosal epithelial cells. According to the method, firstly, the zebrafish is placed in a water body containing antibiotics for fasting culture to solve a problem of microbial self-pollution of zebrafish intestinal tract; then through selection of zebrafish intestinal parts, an intestinal sample that best reflects intestinal function of zebrafish is selected; and finally, through a separation technology of zebrafish intestinal mucosal epithelial cells and setting of culture conditions (primary cell culture medium, CO2 concentration, culture temperature) of cell primary culture, the zebra fish intestinal mucosa epithelial cells with high purity and good growth are obtained. The method provides a theoretical basis for subsequent establishment of the zebrafish intestinal mucosal epithelial cell line, and provides powerful research tools and methods for its extensive use as a fish cell line in genetics, neurobiology, developmental biology, immunology, human disease models, drug screening, toxicology and environmental testing.

The invention discloses a transcription factor C/EBPZ for regulating adipocyte formation and application thereof, belongs to the field of animal molecular genetics and developmental biology, and discloses application of the transcription factor C/EBPZ for regulating and controlling chicken adipocyte formation. The invention discloses an expression mode of chicken C/EBPZ in various tissues and an expression rule of C/EBPZ in a chicken adipose tissue growth and development process by utilizing an RT-PCR technology. According to the invention, an overexpression vector of the transcription factoris constructed, and the C/EBPZ transcription factor is verified to regulate and control the differentiation and proliferation of chicken preadipocytes; the invention further discloses an action mechanism of the transcription factor C/EBPZ in chicken adipose tissue formation. The invention provides a novel application of a C/EBPZ transcription factor in regulation and control of chicken adipocyte formation, and the C/EBPZ transcription factor has practical value in the fields of chicken genetic breeding and animal nutrition.

The invention provides a method for inducing the in-vitro ovulation of mature poultry ovarian follicles. The invention also provides a preparation method of a culturing medium for inducing the in-vitro ovulation of mature poultry ovarian follicles. The preparation method is characterized in that insulin, transferrin, sodium selenite, penicillin, streptomycin, lutropin, follitropin, progesterone, estrogen and an M199 culture medium are mixed under an aseptic condition to prepare a liquid culture medium. Experiments in the invention prove that the induced ovulation of mature ovarian follicles (F1) separated from the ovary of a laying hen (30min after laying) in a laying peak is realized after the in-vitro culturing through using the culture medium provided by the invention. The gonadotropin and steroid hormone in the culture medium provided by the invention approach the gonadotropin and steroid hormone in the poultry in-vivo normal physiological environment, so the culture medium can be widely applied to researches about the poultry ovarian follicle developmental biology.

The invention provides a method for completely silencing a target gene of an embryo by improving microinjection of siRNA. In a developmental biology study, in order to study gene functions, the expression of genes often needs to be inhibited, and siRNA interference is a common means, so that a specific target mRNA can be silenced, the silencing efficiency of the target gent cannot reach 100%, andan expected effect is not achieved. The method for completely silencing the target gene of the embryo by improving microinjection of the siRNA comprises the following steps: S1: removing all granularcells from fertilized oocytes with hyaluronidase; S2: washing the granular cells with a PZM-5 culture medium, placing the washed granular cells into a culture incubator with the temperature of 38.5 DEG C, the humidity of 100% and the carbon dioxide of 5% for recovery for 15min; S3: injecting 15pl of a target liquid with a microinjection pump under an inverted microscope; S4: detecting fluorescencein the embryo after injection under a fluorescence microscope, and determining that the injection is successful if the embryo contains a fluorescent agent. The method is applied to the technical field of developmental biology.

The invention discloses a method for implementing hypothermal anesthesia on fruit flies, and belongs to the technical field of genetics and development biology. Through the method, high-biosecurity and environment-friendly anesthesia on the fruit flies which are import model organisms in the genetics can be achieved, anesthesia is simple in operation, anesthesia time of the fruit flies can be controlled, and the life span and fertility of the anesthetized fruit flies are not affected. The fruit flies are anesthetized at the environment temperature ranging from -15 DEG C to -5 DEG C for 25 seconds to 35 seconds, then the anesthetized fruit flies are transferred to the environment with the temperature ranging from 0 DEG C to 5 DEG C, and follow-up observation and operation of the fruit flies are finished within 20 minutes; eventually, the fruit flies are transferred to the indoor temperature environment, the fruit flies wake, and then follow-up culture and observation of the fruit flies are achieved. The environment temperature is controlled in a programming mode, and when the fruit flies are in the anesthetized state, it can be avoided that ice crystals are formed in the fruit flies due to the physical health due to the fact that the fruit flies are in the low-temperature environment for too long time, and then the life span and the fertility of the fruit flies are damaged.

The invention discloses the method used to directionally induce chicken embryo stem spermatogonium to differentiate into bone cell. Its features are that inoculating the chicken stem spermatogonium to culture bottle without feeder layer; using conditioned medium to induce culturing which is that the DMEM high glucose contains 10% calf blood serum, 2% chicken blood serum, 2mmol/L L-glutamine, 1mmol/L sodium pyruvate, 5.5*10-15ng/ml beta-mercaptoethanol, 10 mul/ml non-essential amino acid, 10-7mol/L dexamethasone, 0.01mol/L beta-phospho glycerol, 0.05g/L vitamin C. The invention has the advantages of advanced technology, simple operation, strong repeatability, providing possible for realizing tissue organ rebirthing and replacing. Thereby hopefully to find out the approach for build cell platform in large scale production, further a new study prospect can be exploited in developmental biology, medicine, genetics fields etc.

The invention relates to a DNA fragment with E2F1 protein binding property containing one or more E2F1 protein binding cassettes, application of the DNA fragment, and a method for detecting transcriptional control activity of E2F1 in cells. By the adoption of the DNA fragment and the method, people can directly detect transcriptional control activity of E2F1 in cells in a targeted mode instead of only detecting the content of transcript or protein of E2F1, so that the function of E2F1 as a transcription factor in developmental biology and pathology development can be analyzed more accurately.

The invention discloses a multiple genotyping kit for detecting the genetic quality of zebra fish. Lines of the zebra fish are diversified; and experimental zebra fish is extremely important for the research on the fields of development biology, genetics, oncology, pharmacology, toxicology, environment friendliness and the like. Particularly, the germplasm degeneration and miscegenation of the zebra fish can seriously affect the scientificity of experimental results, so the kit has a great scientific significance for detecting the genetic quality of the zebra fish. A multiple genotyping scheme kit for a method for detecting the genetic state of the zebra fish is established by taking four lines of the zebra fish, namely TU, TL, AB and WIK as research objects, designing primers and probes for known single nucleotide polymorphism (SNP) sites and utilizing an SNP site primer polymerase chain reaction (PCR) combined ligase detection reaction (LDR) technology.

The invention discloses a method for inhibiting pear seed gibberellin synthetic gene expression, relates to design of complementary chains and cultivation of degreasing cotton wrapped seeds, and belongs to the field of developmental biology and cell biology. According to the method for inhibiting pear seed gibberellin synthetic gene expression by the complementary chains, the influence of lack of gibberellin on seed germination rate can be more effectively learned, and the function of the gibberellin on breaking seed dormancy is more effectively researched. The method is simple, convenient and easy to operate and practically apply and can be widely used for inhibiting other gene expression of different plant seeds.

The invention provides a method for analyzing a biological evolution law through entropy change. The method includes introduction of biomolecule effective volume and free volume concept, definition of an entropy change function, calculation of effective entropy change and free entropy change, analysis of the biological evolution law. Preferably, the method for analyzing the biological evolution law through entropy change also includes introduction of biomolecule effective volume and free volume concept, definition of the entropy change function, calculation of effective entropy change and free entropy change; the biological evolution law can be acquired through calculation of effective entropy change and free entropy change of biological materials, and a new analysis method and theoretical direction are provided for research and development of structural biology, molecular biology, cell biology, developmental biology, genetics and evolutionary biology.

The invention belongs to the field of molecular genetic biology and development biology and discloses an in-vivo gene silencing method taking lentivirus as a tool. The in-vivo gene silencing method taking the lentivirus as the tool comprises the following steps: injecting virus by utilizing a microsyringe; after 2 days, drilling a hole in an air chamber of a chicken embryo and watching and determining the position of an air chamber membrane; inserting the microsyringe into a median membrane of the air chamber to inject the virus. The invention establishes the in-vivo gene silencing method taking the lentivirus as the tool and also successfully establishes a PPARalpha in-vivo silencing chicken embryo model. The method provided by the invention is relatively simple and convenient and operation can be finished under a visual condition; the method is low in cost and high in practicability and the survival rate can reach 80 percent; the transfection efficiency on a heart is high and the in-vivo silencing efficiency can reach about 70 percent. A complicated development signal transduction process is completely kept by in-vivo transfection so that the method can be used as a powerful tool for researching effects of a target gene in development.

The invention discloses a plant seedling culture device, and particularly discloses a water-induction strong-root plant seedling culture device. The water-induction strong-root plant seedling culture device consists of a water holding tray, a net bed bracket, a plant seed holding net bed and absorbent paper, wherein the net bed bracket is placed into the water holding tray; the plant seed holding net bed is placed on the net bed bracket; the absorbent paper is spread on the net bed, the side end of the absorbent paper is in contact with the inner bottom surface of the water holding tray. By adopting the plant seedling culture device, technical support can be provided for selective culture of plant seedling materials which grow robustly, orderly and consistently in the researches of plant developmental biology, physiological plant ecology and the like.

The invention discloses a method for polymerization and in-vitro culture of embryo without zona pellucida before implantation, and belongs to the technical field of embryo engineering and developmental biology. The method comprises the following steps: removing a zona pellucida from an embryo in a 2-8 cell period, putting the embryo into a low-melting-point agarose microdroplet hole covered with embryo-grade mineral oil and a G1 or M16 culture solution for polymerization, and performing in-vitro culture in a tri-gas incubator with 6% CO2, 5% O2 and 89% N2 at 37 DEG C to develop a blastocyst. According to the invention, polymerization and in-vitro culture are respectively carried out on non-zona pellucida fertilized embryos and parthenogenetic embryos of mice before implantation, and it isfound that the blastocyst development rates respectively reach 89.47% and 82.14%, which are obviously higher than those in the prior art, by adopting the gel microdroplet hole method disclosed by theinvention. The method is not only suitable for polyembryos of embryos without zona pellucida before implantation of animals and in-vitro culture of the polyembryos, but also suitable for polyembryos of embryos without zona pellucida before implantation of human beings and in-vitro culture of the polyembryos.

The invention discloses a simple and rapid pig blastaea differential dyeing method and belongs to the technical field of developmental biology. The method comprises the following steps: performing surfactant TritonX100 permeable membrane treatment on the pig blastaea, and performing differential dyeing on trophoderm cells on the outer layer of the blastaea and inner cell mass cells by utilizing the difference of two nucleus dyes in molecular size. According to the method, the experimental steps of performing differential dyeing on the pig blastaea are simplified, the experimental time is greatly shortened, and the experimental cost is reduced.

The invention relates to GATA2 protein binding DNA fragment, which comprises one or more GATA2 protein binding cassettes; the invention also relates to an application of the DNA fragment; and the invention also relates to a method for detecting GATA2 transcription regulating activity in cells. With the application of the DNA fragment and the method provided by the invention, the transcription regulating activity of GATA2 in the cells, instead of merely a transcript or a protein content, can be directly detected in a targeted mode, so that effects of the GATA2, as a transcription factor, in some developmental biology and pathological development can be analyzed more accurately.