30results about How to "Achieve co-cultivation" patented technology

The invention discloses a micro-vessel liver chip based on cell clusters. The micro-vessel liver chip comprises a micro-vessel system located on the upper layer, a blood-vessel-endothelium-like barrier system in the middle and a liver-organ multi-cell co-culture system on the lower layer, wherein the micro-vessel system and the liver-organ multi-cell co-culture system are arranged on own substrates respectively. The micro-vessel system comprises a bent vessel composed of multiple flow-stopping barriers in a staggered mode, and a micro-vessel inlet and a micro-vessel outlet are formed in the two ends of the bent vessel respectively. The blood-vessel-endothelium-like barrier system is composed of a porous film. The liver-organ multi-cell co-culture system comprises a cell cluster enrichment region and a multi-cell co-culture region, and a culture system inlet and a culture system outlet are formed in the two ends of the liver-organ multi-cell co-culture system respectively. A liver disease model can be established, research on pharmacokinetics and medicine activity can be performed, and the micro-vessel liver chip has the advantages of using a small quantity of samples, achieving medicine low consumption and being portable, economical, efficient and accurate.

The invention relates to a novel efficient micro-fluidic multi-cell co-culturing chip and a preparation method thereof. A microstructure and a micro-channel are arranged on the surface of the micro-fluidic chip; the surface of the micro-channel is decorated by bovine serum albumin; under the driving of gravity generated by liquid level difference between a solution inlet and a solution outlet, three cells are implanted in different regions of the micro-channel by using laminar flow phenomenon between liquid flows; finally, the three cells are co-cultured to lay a basis for studies on cell biology; and the novel efficient micro-fluidic multi-cell co-culturing chip provided by the invention is mainly applied to the related field of cell biology, genetics, drug screening and the like. Through adoption of the micro-fluidic chip, the three cells are co-cultured; the micro-fluidic chip has the advantages of intuitive property, convenience, miniaturization and low consumption of reagents and samples; and a brand new cell culturing and analyzing technology is provided for multi-cell co-culturing.

The invention discloses a cell co-culture device. The cell co-culture device comprises a base, a culture medium agitating assembly, a culture dish filled with a culture medium and a plurality of movable cell culture chambers for accommodating cells, wherein the plurality of movable cell culture chambers are movably mounted in the culture dish, permeable membranes allowing non-cells to pass but refusing cells to pass are arranged on the movable cell culture chambers, the movable cell culture chambers are isolated from the culture dish by the permeable membranes, the culture medium agitating assembly is mounted on the base, and a rotating body on the culture medium agitating assembly extends into the culture dish, so that when the culture medium in the culture dish is stirred while the rotating body rotates, cell secretions in the movable cell culture chambers are transferred to corresponding target cells in the movable cell culture chambers. The cell co-culture device can co-culture more cells, avoid cross contamination among different cells, is favorable to observing and harvesting cells, can easily separate specific target cells in the co-culture process for the use of follow-up experiments and can stimulate the growth environment of in-vivo cells.

The invention discloses a tissue-engineered cornea and a preparation method thereof. The tissue-engineered cornea comprises three or more layers of laminated structures, each layer of the laminated structure is a composite layer formed in the mode that a fiber scaffold prepared by electrospinning or electrostatic direct writing or light curing 3D printing is cured and composited with one or more of collagen, an acellular matrix and silk fibroin, the space between every two adjacent layers is inoculated with corneal stromal cells and neurons, the corneal stromal cells and the neurons can grow in the direction of fibers of the scaffold, the corneal stromal cells between every two adjacent layers secrete extracellular matrixes to form a tissue structure similar to the human corneal stroma, and the upper surfaces and the lower surface of the laminated structures are inoculated with corneal epithelial cells and corneal endothelial cells. The tissue-engineered cornea can simulate the corneastructure well, is easy to prepare and good in biocompatibility and is easily fused with tissue.

The invention belongs to the field of aquaculture and discloses a compound bacteria preparation and a preparation method and application thereof. The preparation method includes: (1), preparing seeds;(2), inoculating bacillus coagulans; (3), inoculating clostridium butyricum; (4), inoculating lactobacillus and saccaromycetes mixed seed liquid to obtain the compound bacteria preparation. The compound bacteria preparation can be applied to aquaculture and can effectively degrade nitrite in an aquaculture pond, purify water and promote healthy culture and efficient production of aquaculture industry of China.

The invention relates to a microfluidic chip for researching the interaction between cells and the preparation method thereof. A microstructure and a micro channel are arranged on the surface of the microfluidic chip. The two types of cells are fixed in respective cell culture areas respectively. The cells interact with each other through the micro channel. The interaction between the two types of cells during the process of growth can be visually researched with a microscope, as a result, a new thought and a new research technique for researching cell biology are provided; and moreover, the microfluidic chip is mainly applicable to fields relevant to cell biology, genetics, drug screening and the like. The microfluidic chip achieves co-culture of two types of cells, has the characteristics of visuality, convenience, miniaturization, less dosage of agent and fewer samples, thereby providing a novel analyzing technology for co-culture of two types of cells.

The invention relates to a method for amplification of hemopoietic stem cells. The method comprises the steps of separation of mononuclear cells, preparation and culture of microencapsulated mononuclear cells, and amplification of hemopoietic stem cells. The method can realize the amplification of hemopoietic stem cell as well as carries out culture on a large scale, and can play a significant role in blood biology and clinic application and research.

The invention discloses a multi-cell co-culture model based on fiber hydrogel and a preparation method of the model. The multi-cell co-culture model comprises a multi-layer oriented nanofiber membraneand GelMA hydrogel which is poured into the multi-layer oriented nanofiber membrane to form gel and is of a three-dimensional porous structure. The multi-layer oriented nanofiber membrane can be prepared by utilizing an electrostatic spinning technology. By using the multi-cell co-culture model, a cell is inoculated on the multi-layer directional nanofiber membrane, a second cell is inoculated onthe upper surface of a fiber hydrogel stent, and a third cell is inoculated below the fiber hydrogel stent, so that in-vitro co-culture of various cells is realized. The multi-cell co-culture model not only can realize co-culture of multiple cells, but also has good mechanical properties and high bionic properties, and can provide structures and properties of extracellular matrixes capable of simulating natural tissues for the multiple cultured cells.

The invention relates to a micro-fluidic cell culturing chip and a preparation method thereof. A microstructure and a micro-channel are arranged on the surface of a micro-fluidic chip; two cells are respectively fixed in specific cell culturing regions; signal transmission and interaction between the cells are carried out between the cells through the micro-channel; under a microscope, the interaction between the two cells in the growing process can be intuitively studied and metabolites in the cell growing process are analyzed; a new idea and a new research technique are provided for researches on the cell biology, and the micro-fluidic cell culturing chip is mainly applied to the related field of cell biology, genetics, drug screening and the like. Through adoption of the micro-fluidic chip, co-culturing of the two cells is achieved, the operation is simple, parallel implantation and co-culturing of the two cells are realized, the usage amounts of reagents and samples are reduced, the cell implanting process is simplified, the interaction between the two cells in the growing process can be intuitively studied under the microscope, the metabolites in the cell growing process is analyzed and a brand new analysis technology is provided for researches on the cell biology and the micro-fluidic cell culturing chip provided by the invention is portable, economic, rapid, efficient and accurate.

The invention relates to a direct contact type cell co-culture device which belongs to the technical field of cell co-culture devices. The direct contact type cell co-culture device comprises a culture hole, a culture plate and an upper cover, wherein the culture hole is arranged on the culture plate, and the upper cover is covered on the culture plate and seals the culture hole; an isolation plate is arranged in the culture hole, and the culture hole is divided into a plurality of small culture chambers. A reverse hook is arranged on the outer side wall of the upper part of the culture hole, two upright edges of the isolation plate are installed in a side fixing groove arranged in the culture hole, the bottom edge of the isolation plate is provided with an elastic pad, and the elastic pad is pressed on the bottom surface of the culture hole; when the culture hole is not pressed, the height of the isolation plate is slightly higher than the depth of the culture hole, and the upper edge of the isolation plate is installed in an upper fixing groove; and the end of the upper fixing groove is provided with a connecting line with a circular ring. Before being unsealed, the absolutely aseptic state of the cell culture device can be ensured. The invention has the advantages of simple operation and reliable effect, can clearly observe and contrast the situation of the growth change of cells at the edges of the small culture chambers, can realize the co-culture of various cells and does not influence the normal adherence and multiplication of the cells.

The present invention provides establishment and applications of a three-dimensional liver model based on qualitative filtration paper. The three-dimensional liver model is mainly based on the normalphysiological structure characteristics of liver, wherein liver cells hiHep obtained through transdifferentiation are directly induced with fibroblasts, a traditional primary liver cell or liver cellline monolayer culture model is improved, and qualitative filtration paper as a scaffold material for cell culture is placed in a 24-well plate, and is suspended in a culture solution, such that the complete material transfer and nutrient component exchange can be performed so as to achieve the three-dimensional growth of cells on the filtration paper. According to the present invention, the co-culture of hiHep cells and umbilical vein endothelial cells (HUVECs) is performed on the qualitative filtration paper to obtain the liver model based on the filtration paper, such that the three-dimensional liver tissue capable of being subjected to long-term in vitro culture and maintaining the specific functions of liver is achieved; with the obtained three-dimensional liver tissue, the in vitro test and evaluation can be performed on a series of hepatotoxicity drugs; and the obtained three-dimensional liver tissue is mainly used for the in vitro research of the long-term toxicity of drug molecules to liver tissues and the liver function injury process.

The invention discloses a laboratory microbial culture device. The device comprises a culture box; the inner walls of culture rooms are fixedly connected with heat preservation layers; the upper surfaces of rollers are fixedly connected with culture apparatuses; culture dishes are arranged in grooves; the inner walls of the culture rooms are fixedly connected with heating plates; a temperature sensor is arranged on one side of each heating plate; an oxygen concentration detector and a humidity detector are fixedly connected to the other inner side wall of each culture room; and a water tank and an air disinfection tank are fixedly connected to the back face of the culture box. According to the laboratory microbial culture device, through the arrangement of the four culture rooms, flow dividing pipes and water dividing pipes, the purposes of co-culture of various microorganisms and improvement of the use function are achieved; through the arrangement of the temperature sensors, the humidity detectors and the oxygen concentration detectors, the purpose of detecting and adjusting the temperature and the humidity is achieved; and through the arrangement of the air disinfection box andan ultraviolet lamp, the purpose of preventing external microorganisms from entering the culture rooms is achieved.

The invention provides a high-throughput microorganism isolation culture method, which comprises the following steps: performing primary embedding on microorganisms to be isolated by taking agarose as an embedding material, performing secondary embedding to obtain microspheres by taking sodium alginate as an embedding material, and isolating the microspheres by virtue of a flow cytometer. According to the method, an in-situ sample can be directly added into a double-layer sphere culture system to implement co-culture of the microspheres and the in-situ sample. In such a co-culture manner, the difficult-to-culture microorganisms can be cultured more easily, and samples, particularly precious samples which are difficult to process such as nodules, can be fully utilized to increase the utilization rate of the precious samples.

The invention discloses collagen-based self-healing hydrogel and a preparation method thereof. The method comprises the following steps: uniformly mixing a collagen solution and a mixed solution of guar gum and dialdehyde guar gum, adding a borax solution, stirring and mixing, and performing vacuum defoaming, so as to obtain the collagen-based self-healing hydrogel. A feasible approach and a novelthought are provided for the preparation of the collagen-based self-healing hydrogel through the scheme. In addition, the self-healing hydrogel can properly meet the requirement of injectable hydrogel, and structures damaged during injection can be reconstructed in organisms; moreover, the self-healing hydrogel as a three-dimensional cell culture carrier has unique superiorities, can properly simulate cell living environment, is beneficial to transfer of contact information among cells and to realizing of coculture of various and multi-layer cells.

The invention discloses an in-vivo heart organ chip and method. The in-vivo heart organ chip comprises a substrate device and a culture solution perfusion device, wherein a PDMS thin film is arranged on a surface of the substrate device, and the culture solution perfusion device is arranged at a top of the PDMS thin film to simulate blood flowing condition; the substrate device comprises a substrate, a PDMS mold is arranged on a surface of the substrate, and the PDMS mold is provided with a micro gas channel; the PDMS thin film comprises a suspended thin film, and the suspended thin film is provided with a strain sensing part, an induction electrode and a stimulating electrode.

The invention provides a microfluidic platform, a preparation method and application thereof. The microfluidic platform comprises a plurality of culture chambers, the culture chambers are connected through micro-channels, the depth of the micro-channels is determined through axons of cells or tissue in the culture chambers, so that the axons pass through the micro-channels, and the cells and tissues corresponding to the axons cannot pass through the micro-channels; and the cells or tissues in different culture chambers are connected through axons. According to the invention, physical and fluid isolation is carried out on damaged nerve cells or tissues to realize co-culture.

The invention relates to a pit mud flora enrichment method based on oligoculture and application thereof. The pit mud flora enrichment method based on oligoculture comprises the following steps of: inoculating pit mud into an enrichment medium for culture to enrich pit mud flora; wherein the enrichment medium comprises a basic medium and a selective carbon source, the basic medium comprises mineral element substances, trace element substances, B vitamins, yeast powder, peptone, sodium bicarbonate, sodium sulfide and water, the selective carbon source comprises short-chain fatty acid and / or short-chain fatty acid salt. The enrichment method can perform culturable enrichment on various uncultured flora, can be applied to preparation of methane bacteria or butyric acid utilization flora bacterial liquid, methane flora-butyric acid utilization flora co-culture, butyric acid utilization bacteria or methane bacteria activity detection, artificial pit mud manufacturing and maintenance and thelike, and provides a leading technology for pit mud functional microorganism resource mining.

The invention relates to a 3D pseudo-skin construction method based on organs-on-chips and directional differentiation of induced pluripotent stem cells. According to the method, a 3D spinning supportis placed in sterilized organs-on-chips, fibroblast is inoculated on one surface of the 3D spinning support, keratinocyte is inoculated on the other surface of the 3D spinning support, then two culture mediums (CnT-07 epidermal keratinocyte medium and DMEM with 10 vol.% of FBS and 1 vol.% of penicillin-streptomycin) are introduced into the organs-on-chips to culture fibroblast and keratinocyte respectively, and the co-culture of fibroblast and keratinocyte and cell proliferation are realized. The construction method utilizes a stem cell differentiation technology to obtain required cells, thecell inoculation is performed on a nano structure, fibroblast and keratinocyte are co-cultured in organs-on-chips, and the method is used for epidermal tissue repair, is used to establish a drug model, and is the hot spot of research at present.

The invention discloses a device and a method for arranging a plurality of cells at the same plane and controlling the cells, comprising: a thin film with micro-hole arrays which is located at the upper surface of a base; a PDMS stamp whose lower surface is provided with a micro-groove unit and is coated on the thin film; the micro-groove unit comprises: an intermediate groove and a left-side groove and a right-side groove which are located at the left side and the right side of the intermediate groove; the middle sections of the left-side groove and the right-side groove are parallel to the intermediate groove; the both end sections except for the middle sections of the left-side groove and the right-side groove slant towards the direction away from the intermediate groove; the end of the groove is provided with a vertical through hole which is communicated with the corresponding groove; the holes of the thin film are all located in the micro-groove of the stamp; and a micro-pipelineis provided with different cell suspensions, the cell grows in the micro-pipeline and adheres to the micro-pipeline; the stamp is removed to realize the co-culture of a plurality of cells at the sameplane; and then the thin film is removed to realize the free movement of the plurality of co-cultured cells at the same plane; the method is simple to operate, the used material is cheap, and the invention can be used for high-throughput screening of useful drugs for cells.

A tissue-engineered cornea and a preparation method thereof, the tissue-engineered cornea comprises a laminated structure of at least three layers, wherein each layer is a fiber scaffold prepared by electrospinning or electrostatic direct writing or photocuring 3D printing, collagen, decellularized matrix, One or more kinds of silk fibroin are solidified and combined to form a composite layer, and corneal stromal cells and neurons are seeded between adjacent two layers, and the corneal stromal cells and neurons can be along the direction of the fibers of the scaffold The corneal stromal cells between two adjacent layers secrete extracellular matrix to form a tissue structure similar to human corneal stroma. The upper and lower surfaces of the laminated structure are inoculated with corneal epithelial cells and corneal endothelial cells respectively. The tissue engineered cornea can well simulate the structure of the cornea, is simple to prepare, has good biocompatibility, and is easy to fuse with tissue.

The invention discloses a micro-vessel liver chip based on cell clusters. The micro-vessel liver chip comprises a micro-vessel system located on the upper layer, a blood-vessel-endothelium-like barrier system in the middle and a liver-organ multi-cell co-culture system on the lower layer, wherein the micro-vessel system and the liver-organ multi-cell co-culture system are arranged on own substrates respectively. The micro-vessel system comprises a bent vessel composed of multiple flow-stopping barriers in a staggered mode, and a micro-vessel inlet and a micro-vessel outlet are formed in the two ends of the bent vessel respectively. The blood-vessel-endothelium-like barrier system is composed of a porous film. The liver-organ multi-cell co-culture system comprises a cell cluster enrichment region and a multi-cell co-culture region, and a culture system inlet and a culture system outlet are formed in the two ends of the liver-organ multi-cell co-culture system respectively. A liver disease model can be established, research on pharmacokinetics and medicine activity can be performed, and the micro-vessel liver chip has the advantages of using a small quantity of samples, achieving medicine low consumption and being portable, economical, efficient and accurate.

The invention discloses a preparation method of a heterogeneity 3D tumor composite multicellular sphere invasion model, and mainly solves the problems of high model preparation cost and high technical operation difficulty in the prior art. The method comprises the following steps of: 1) obtaining a single-cell suspension; 2) forming initial multiple tumor cells; 3) establishing a tumor cell strain capable of stably expressing a GFP fluorescent protein; 4) forming tumor composite multicellular cells; and 5) constructing the heterogeneity 3D tumor composite multicellular sphere invasion model. The invention successfully develops a simple, efficient and repeatable method for establishing the heterogeneity 3D tumor composite multicellular sphere invasion model, has a better condition for large-scale popularization, and also provides an important research model for the fields of research on tumor heterogeneity, tumor microenvironment, tumor immunity, tumor drug screening and the like, and the requirements of scientific research and clinical application are fully met.

The invention relates to a method for constructing a 3D pseudo-epidermis based on organ chips and directed differentiation of induced pluripotent stem cells, comprising the following steps: placing a 3D spinning scaffold inoculated with fibroblasts on one side and keratinocytes on the other side in an inert environment. Into the organ chip after bacteria, two kinds of media, CnT-07 epidermal keratinocyte medium and DMEM (containing 10% volume ratio FBS, 1% volume ratio penicillin-streptomycin), were passed through to culture keratinocytes and fibroblasts respectively , to achieve co-culture and cell proliferation of keratinocytes and fibroblasts. In the construction method of the present invention, the required cells are obtained by using the stem cell differentiation technology, the cells are inoculated on the nanostructure, and the co-cultivation of keratinocytes and fibroblasts is realized in the organ chip, so that the repair of epidermal tissue has become a current research hotspot. and new methods for drug modeling.

The invention discloses a micro-fluidic chip as well as a construction method and application thereof. The micro-fluidic chip comprises a polydimethylsiloxane layer and a substrate layer, wherein the polydimethylsiloxane layer comprises three or more than three micro-channels; the micro-channels are connected through a plurality of micro-channels which are arranged in parallel; the micro-channel is provided with a pore channel connected with the outside; the substrate layer is at least one of a glass substrate, a polydimethylsiloxane substrate or a polystyrene substrate, the preparation method of the micro-fluidic chip is simple, and the culture of tumor nerve cells can be realized. A pancreatic cancer-nerve-stromal cell microenvironment model constructed by utilizing the micro-fluidic chip disclosed by the invention can realize joint construction of a pancreatic cancer stromal microenvironment and a neural microenvironment, and can also perform qualitative and quantitative evaluation on nano-drugs targeting the neural microenvironment and the stromal microenvironment; the method plays an important role in screening related drugs for treating pancreatic cancer.

The invention provides a high-throughput microorganism isolation culture method, which comprises the following steps: performing primary embedding on microorganisms to be isolated by taking agarose as an embedding material, performing secondary embedding to obtain microspheres by taking sodium alginate as an embedding material, and isolating the microspheres by virtue of a flow cytometer. According to the method, an in-situ sample can be directly added into a double-layer sphere culture system to implement co-culture of the microspheres and the in-situ sample. In such a co-culture manner, the difficult-to-culture microorganisms can be cultured more easily, and samples, particularly precious samples which are difficult to process such as nodules, can be fully utilized to increase the utilization rate of the precious samples.