60 results about "Cell Separation Technology" patented technology

The invention relates to a kit for processing human marrow, cord blood and peripheral blood stem cells and a stem cell processing method. The kit radically solves the problems of high manufacture cost, low cell activity, indefinite reaction of markers in a human body, mobilizing agent injection causing patient pain, long obtainment time, complication, clinical unsuitability, and the like in the prior cell separation technology. The invention has the technical scheme that the kit comprises the following three reagents: (1) a diluent comprising 0.9 percent of sodium chloride injection or PBS solution, (2) a precipitator comprising 6 percent of hydroxyethyl starch or 0.2-1 percent of methyl cellulose and (3) a separating solution which has the density of 1.074-1.076 and is prepared by saccharosan and diatrizoate. The kit is easy to store and transport and convenient and rapid to use and can be produced in industrialization.

The invention belongs to the technical field of cell separation, and particularly discloses an automatic medical cell separation device which comprises a separation tube. The separation tube is internally provided with a transmission rod, the transmission rod is provided with a structure which can extrude and crush or smash a medical cell, the top end of the transmission rod is connected with a driving motor, and an outlet is arranged on the bottom part of the separation tube and is provided with a plug cover. By the medical cell separation device, a large cell tissue can be conveniently and easily extruded and crushed or smashed into cell particles, the operation process of cell separation of the separation device and an operation can be simultaneously carried out in an operating room, nonnutritive time after the medical cell separation can be effectively shortened, pollution opportunities and harms of the cell can be reduced, and the quality control of the cell treatment can be convenient; simultaneously, the storage and transportation of samples are greatly convenient, the standard operation can be carried out, and the repeatability of a research can be increased; and automation control and personalized customization can be carried out according to different tissue cells, so that the use is simple and convenient.

The invention discloses a humanized anti-H7N9 avian influenza virus high-affinity antibody 10K filtered and obtained based on a single cell separation technology, the amino acid sequences of light chain and heavy chain variable regions of the antibody are shown as SEQ ID No. 2 and SEQ ID No. 5 respectively. The high-affinity specificity of the antibody is combined with H7N9 avian influenza virus 7 type hemagglutinin protein, and can mediate the kill and wound (ADCC) of effector cells using NK cells as main parts for H7N9 influenza virus infected cells. The antibody 10K can be used for therapeutic development of highly pathogenic avian influenza infection, and also can be used for development of H7N9 influenza virus antigen dectection reagents.

The invention discloses a kit for treating human bone marrow, umbilical cord blood, and peripheral blood cells, and provides a kit for treating human bone marrow, umbilical cord blood, and peripheral blood cells and a cell treatment method which are strong in operationality, high in clinical safety, and convenient for clinical popularization. The kit comprises four reagents: No. A liquid is a diluent; No. B liquid is density fluid; No. C liquid is washing liquid; No. D liquid is erythrocyte-removing liquid. The invention fundamentally solves the problems of high cost, low cell activity, undefined human body influence for markers entering human body, pain for patients due to mobilization agent injection, cumbersome and inapplicable operations, and the like for current cell separation technology.

The invention relates to the technical field of cell culture and cell separation and in particular discloses a preparation method for improving the yield of umbilical cord derived mesenchymal stem cell primary cells. On the basis of culturing and separating mesenchymal stem cells on a traditional Wharton's jelly tissue block in a disposable adherence manner, Wharton's jelly tissue blocks are finely collected at different time of a later period of culture and are repeatedly adhered; P0 generation mesenchymal stem cells can be continuously collected at different time. According to the method provided by the invention, the utilization rate of Wharton's jelly is maximized through a property that the mesenchymal stem cells continuously move out from Wharton's jelly tissues to grow; compared with Wharton's jelly tissue block disposable adherence culture, the obtained quantity of the P0 generation mesenchymal stem cells can be increased by two times at least; the culture density of the mesenchymal stem cells, the concentration of serum replacements in a culture medium and the culture time are subsequently controlled; when the cells are sub-cultured to a P3 generation, the obtained cell quantity can completely meet the requirements of cell storage and clinical transplantation.

The invention belongs to the technical field of cell separation, and particularly relates to a separation method for islet cells of a mouse. The method comprises the following steps: narcotizing the mouse, disinfecting the skin and cutting along the center of the abdomen to fully expose the pancreas; shearing the ligament organization connected with the pancreas of the spleen, clamping to close the junction of the splenic vein and the spleen and cutting the heart broken, inserting a needle at the intersection of the splenic vein and the portal vein to the splenic vein, and injecting collagenase filling liquid; immersing the filled pancreas of the spleen into the collagenase filling liquid after the pancreas of the spleen is dissociated from the abdominal cavity; performing water bath slaking on the pancreas collagenase solution, performing centrifugal separation after adding Hank's liquid and taking the sediment; and performing heavy-suspending sedimentation, filtration and centrifugal treatment on the sediment to obtain the islet cells of the mouse. The separation and purification method for the islet cells of the mouse, provided by the invention, is simple and feasible in operation, and the separated islet cells have no obvious differences with those obtained by a CBD method in forms and functions, so that the selectable technologies for separation of pancreas islet of the mouse are enriched, and the separative power is improved.

The invention discloses a preparation method of an HLA-A0201 (Human Leukocyte Antigen-A0201) restrictive anti-MAGE (Micro Array and Gene Expression) antigenic specificity CTL (Cytotoxic T Lymphocyte). The method comprises the following steps of: carrying out hemapheresis to collect peripheral blood mononuclear cells; enriching and purifying CD8+T lymphocytes; utilizing a mature dendritic cell loaded with an HLA-A0201 restrictive anti-MAGE antigen polypeptide to stimulate the CD8+T lymphocytes; utilizing rhIL-2 and rhIL-7 to accelerate the T cells to grow; utilizing a Tetramer marking method and a flow type cell separation technology to purify the target CTL; utilizing an anti-human CD3 monoclonal antibody covered by a solid phase and an IL-2 to stimulate the target CTL to grow; adding an autologous PBMC (Peripheral Blood Mononuclear Cell) radiated by gamma rays to enhance the activation of the target CTL; adding rhIL-15 to cultivate and reproduce; and collecting and identifying. The prepared target CTL has high purity, high reproduction capability, high killing activity and high ratio CTL-CM, and can be used for immune treatment of tumors and the like.

The invention relates provides a batch preparation method of high-purity salvianolic acid A based on physical and chemical properties and chromatograph behavior of a chemical component in salvia miltiorrhiza as well as adaptability of salvia miltiorrhiza to a cell separation technology. In the method, a salvianolic acid B transformation liquid containing salvianolic acid A is subjected to three-time chromatograph purification, namely hyphenated chromatography, reversed-phase chromatography and normal-phase silica gel chromatography, wherein the hyphenated chromatography refers to that macroporous resin frontal chromatography and macroporous resin displacement chromatography are jointly used. The product prepared by the method is controllable in quality, is acceptable in cost and can be used for injection medicaments. The content of salvianolic acid A prepared by the method is more than or equal to 98%, the total recovery of salvianolic acid A is more than or equal to 10% (based on salvianolic acid B), and the total yield of salvianolic acid A is more than or equal to 0.5% (based on salvianolic acid B).

The invention discloses a cell sample extraction and subpackaging device, and relates to the technical field of cell separation. The cell sample extraction and subpackaging device comprises a supporting frame, wherein a centrifugal mechanism used for driving a rotating groove to rotate is in driving connection to a driving motor II; a vertically-arranged rotating column is rotationally installed on the supporting frame; and a subpackaging mechanism used for subpackaging cell samples is installed at the lower end of the rotating column. According to the cell sample extraction and subpackaging device, the arranged centrifugal mechanism can drive the rotating groove to drive a centrifugal groove to rotate, and the centrifugal separation effect of cells is achieved; an arranged suction head can convey the cells in the centrifugal groove into a storage box under the effect of a conveying mechanism, the suction head can be driven by a lifting mechanism to vertically move, the cells at different depth positions in the centrifugal groove are extracted, and cell extraction operation and adjustment are flexible and efficient; and the arranged subpackaging mechanism can stably and rapidly subpackage the extracted cells, the effect of storing and loading the cell samples on a large scale is achieved through a plurality of sets of test tubes on a rotating disc, and a large number of cell samples can be conveniently used in biological experiments.

The invention provides a cell separation technology, and particularly relates to a separation method of macrophage in a thyroid papillary carcinoma tissue. The separation method comprises the following separation steps of: placing the collected thyroid papillary carcinoma tissue into D-hanks liquid; flushing; removing D-hanks liquid and connective tissues; cutting the collected thyroid papillary carcinoma tissue into blocks; adding II type collagenase into the blocked tissue; performing digestion reaction for 1 to 3 hours at temperature of 35 to 40 DEG C; adding an 1640 culture medium containing 10% of fetal calf serum for stopping the digestion reaction; filtering; centrifuging; removing supernatant; resuspending and planking cell sap through the 1640 culture medium; transferring into a cell incubator containing 5% of CO2 for incubating; fully washing through PBS (Phosphate Buffer Solution); re-culturing the obtained anchorage-dependent cells; collecting the supernatant; and freezing and storing. According to the separation method, the separated macrophage is circular and can be tightly adhered to a petri dish and shows positive by staining by CD68; the positive rate reaches more than 95%, namely, the macrophage is separated successfully.

A plant-protein product and the method for preparing the same are provided. The plant-protein product is obtained from a plant-protein raw material which is treated by a single-cell separation technique, and the method for preparing the plant-protein product comprises steps of providing the plant-protein raw material; and treating the plant-protein raw material by the single-cell separation technique to prepare the plant-protein product.

The invention discloses a method for preparing animal mature neuron single cells, and belongs to the technical field of separation of mature neuron single cells. According to the method, firstly, D-PBS is utilized for conducting perfusion on a mouse, blood cells are removed, and the influence of the blood cells on subsequent separation is avoided; Hibernate-A is utilized as a culture medium in thedigestion process, without considering the problem of early oxygen introduction or oxygen introduction in the digestion process, the time can be effectively saved by at least 30 minutes, and the damage to neurons is reduced; by means of the method, the single cells can be prepared without special instruments; through the method, oligodendrocytes, astrocytes and neurons can be simultaneously separated out, sequencing or function analysis can be conducted on different types of nerve cells in the same batch of samples, the experimental errors can be reduced, and the cost is reduced.

The invention belongs to the technical field of cell separation and particularly discloses a separation culture method for pig intestinal stem cells. The separation culture method comprises following steps: firstly screening specific antibodies applicable to the separation of the pig intestinal stem cells, wherein the antibodies include CD24, CD44 and CD166, and the antibody CD24 is derived from ML5 monoclone; screening the pig intestinal stem cells in a targeted manner by virtue of the antibodies and a flow cytometry, establishing a three-dimensional culture system, and culturing intestine-like groups. According to the separation culture method, good materials are provided for the regeneration study of epithelia of intestinal tracts and tissue specific stem cells transplantation.

The invention belongs to the technical field of single cell separation and specifically relates to a source component and a device for inducing a high-voltage impulse wave to separate single cells through laser photolysis. In the source component provided by the invention, the original cell culture layer is designed as a micro-array pore structure and a cell culture fluid fills the through holes on the micro-array pore structure, so that the cells in the culture fluid are firstly pre-separated, the situation of one sputtering liquid drop containing a plurality of cells is avoided and the cell transferring precision and the separation efficiency are increased. According to the invention, the source component and a packaging component are assembled into a packaged structure, so that a sealed technological environment is formed in a single cell separation area, the suitable air humidity is guaranteed, the cell inactivation caused by dry air is prevented and the excellent cell survival environment is maintained.

The invention relates to the technical field of cell separation and particularly discloses a separating and extracting method for primary liver parenchyma cells and application of the primary liver parenchyma cells. The method disclosed by the invention comprises the steps: injecting perfusate I and perfusate II into a hepatic tissue mass to digest and separate the liver parenchyma cells so as toobtain a cell suspension, then, subjecting filter liquor, obtained after filtering the cell suspension, to normal-temperature centrifugation for 5 times sequentially, wherein centrifugation conditionssequentially are 1000-1580rpm / min*3-5min, 500-650rpm / min*3-5min, 200-300rpm / min*4min, 200-300rpm / min*4min and 200-300rpm / min*4min. According to the method, the operation is simple, consumed time is short, low-temperature centrifugation equipment is not required, red blood cells and cell debris can be efficiently removed, and obtained liver parenchyma cells are relatively high in survival rate andpurity.

The invention discloses a hepatic stellate cell separation and preparation method. The hepatic stellate cell is prepared through the steps of perfusion, stripping, and density gradient separation. The perfusion is carried out successively with a perfusion solution I, a perfusion solution II and a perfusion solution III, wherein the perfusion solution I comprises a buffer solution I; the perfusion solution II is prepared by adding 0.22-0.38 mg of pronase E to 1 ml of a buffer solution II; and the perfusion solution II is prepared by adding 0.52-0.68 mg of collagenase D to 1 ml of the buffer solution II. The invention belongs to the technical field of cell separation, wherein cell surface substrates are completely enzymolyzed successively by means of single enzyme and then compound enzymes matched with the buffer solution II, so that yield of the hepatic stellate cells are increased.

The invention relates to a cells separating technology, and provides a tumor infiltrating lymphocytes separation method. The method is characterized by adding tumor tissue masses in a trypsin-EDTA solution for immersion and incubation, fully digesting and dispersing the tumor tissue masses; separating and collecting single cell and being resuspended by the trypsin-EDTA solution for multitime density gradient centrifugation, incubating and separating by an erythrocyte lysate, using calf serum-containing PBS buffer or resuspending to obtain the dissolved tumor infiltrating lymphocytes. In the invention, an enzymatic digestion method and a machinery method are combined for dual separating the enzymatic digestion with high efficiency, compared with the machinery method or the enzymatic digestion method, the survival rate of the obtained tumor infiltrating lymphocytes is high, and the method is simple and easy to operate. The obtained tumor infiltrating lymphocytes can be massively propagated through in vitro stimulation of interleukin 2 (1L-2), have antineoplastic activity with specialty and high efficiency, and provides powerful basis and new approach for treating tumour.

The invention relates to a cell separation technology, belongs to the field of biomedicine, and specifically relates to a cell separation method based on laser array coding and photo-induced micro-bubble precipitation. In prior art, in the fields like biosensor, mankind functional genome carrier research, rare cell screening, judicial evidence obtaining of sex crimes, and so on, the target cells for rapid separation and detection comprise too much information, the provided method solves the problem mentioned above. The matured image identifying technology and automatic control technology are effectively combined in the method, and thus the technologies are not just confined in single micro fluidic array or applied to single purpose.

The invention relates to the technical field of cell separation, in particular to adipose tissue digestive juice and a method for rapidly obtaining an SVF (stromal vascular fraction) cell. The adiposetissue digestive juice comprises the following ingredients: I-type collagenase, HEPES buffer solution and a DEMEM / F12 culture medium. According to the adipose tissue digestive juice provided by the invention, the pH value of a digestive system can be stably maintained between 6.8-8.2, so that the activity of the I-type collagenase is optimal. Therefore, complete digestion can be realized in a short time. The time for obtaining the SVF according to the traditional digestive scheme is 40 min, while the SVF with high quality and high quantity can be obtained within 15-20 min in such a way that the digestive juice is adopted for digesting adipose tissues.

The invention relates to the technical field of vascular cell separation, in particular to a method for separating and enriching vascular endothelial progenitor cells from peripheral blood. The method comprises the steps of obtaining a peripheral blood sample, and using PBS phosphate buffer for dilution; adding diluted blood into a hydroxyethyl starch solution, performing centrifuging and sucking monocular cell sap of a middle albuginea cell layer; using the PBS phosphate buffer for dilution, performing centrifuging, abandoning a supernatant, and performing resuspension on residues through a nutrition solution; performing culture transferring. The method is high in separation efficiency, adopts few parenchyma cells, only needs a small number of experiment instruments, is good in separation effect and has the repeatability.

The invention relates to the technical field of cell separation, in particular to a separate culture method of primary tumor cells. A composite enzyme solution is adopted for decomposing cancer tissue, wherein collagenase, trypsin and hyaluronidase in the composite enzyme solution mutually cooperate, looseness of bridging structures among the cells can be accelerated, the cancer tissue is converted to be flocculent from being blocky, thus, the cells in cell aggregate are evenly dispersed in cell suspension, and the numbers of cell alliances and single cells suspended in cell sap are increased;after inoculated culture is completed, the cells easily adhere to walls to grow, and therefore the survival rate of the cells is increased; serum extracted from the body of a patient suffering from tumors is added into a culture medium, a microenvironment most suitable for the growth of the cancer cells is provided for the cancer cells, the culture period is shortened, the culture success rate is80% or above, and the clinic consistency rate is 95% or above.

The invention relates to the technical field of cell separation, in particular to a centrifugal separation device for cell separation. The device comprises a base, a controller and a cell homogenizing tank are fixedly mounted on the base, a centrifugal barrel is rotatably mounted at one end of the cell homogenizing tank, a centrifugal cavity and a precipitation cavity are respectively arranged in the centrifugal barrel from top to bottom, a stepping motor used for driving the centrifugal barrel is fixedly installed on one side of the cell homogenizing tank, the output end of the stepping motor is connected with the centrifugal barrel through a transmission part, a transposition disc is rotationally installed on the base, a plurality of material storage containers are detachably installed on the transposition disc in the circumferential direction, a material receiving disc is arranged over the transposition disc, and plurality of falling grooves are formed in the material receiving disc in the circumferential direction. By arranging a linkage structure, the automation degree of the whole process is improved, and the period of the whole separation procedure is shortened.

The invention provides a method for purifying salvianolic acid A by using reversed-phase chromatography based on physical and chemical properties and chromatograph behavior of a chemical component in salvia miltiorrhiza as well as adaptability of salvia miltiorrhiza to a cell separation technology. In the method, a used reversed-phase chromatography filling material is reversed-phase resin CG161, SBC-MCI-GEL or MCI-GEL-CHP-20P, and the weight ratio of salvianolic acid A to used the reversed-phase chromatography filling material is 1: (30-50); and the reversed-phase chromatography is operated by the following steps: sampling a crude salvianolic acid A aqueous solution; eluting with a 20%-25% alcohol solution until the salvianolic acid A is detected in eluent; eluting with a 35%-40% alcohol solution and collecting salvianolic acid A until the content of salvianolic acid A in eluent is minimum; and finally, eluting with a 50-70% alcohol solution and collecting salvianolic acid A containing impurities. The product prepared by the method is stable and controllable in quality and is acceptable in cost.

The invention provides a preparation method of a separation tube for separating tumor cells, and belongs to the technical field of cell separation. The preparation method comprises the following stepsof firstly adding separation gel to the separation tube, performing centrifuging so as to enable the separation gel to sink to the bottom of the tube, then adding separation mediums, then slowly adding body cavity hydrops or anticoagulation whole blood to an upper layer of the separation mediums, performing centrifuging at room temperature, firstly sucking and discarding liquid at the topmost surface of the separation tube so as to remove cell debris, then dumping residual liquid on the separation gel layer into a collecting tube, then performing centrifuging at room temperature, performing cell re-suspending with PBS so as to obtain the tumor cells, wherein the density of the separation gel is 1.06-1.08g / mL, and the density of the separation mediums is 1.05-1.06g / mL. The preparation method disclosed by the invention can be used for separating and circulating the tumor cells from the tumor cells in the body cavity hydrops, the preparation method is simple, convenient and efficient, and the purity of the separated tumor cells is high.

The invention discloses a cell separator with high separation efficiency. The cell separator comprises a mounting box, the top of the mounting box is slidably connected with a primary filtering mechanism, the primary filtering mechanism comprises a filtering box, the two sides of the filtering box are fixedly connected with fixing plates, the bottoms of the two fixing plates are slidably connected with the top of the mounting box through guide rails, the bottoms of the fixing plates are fixedly connected with tooth plates, the top of the mounting box is fixedly connected with first driving mechanisms, the bottom of the mounting box is fixedly connected with a filtering frame through a second driving mechanism, and the second driving mechanism comprises telescopic rods.The invention relates to the technical field of cell separation. According to the cell separator with the high separation efficiency, tissue fluid is filtered twice through the filtering box and the filtering frame, stem cells can be automatically screened out, the workload of workers is relieved, the filtering frame can move up and down in the mounting box, the filtering speed is increased, the working efficiency is improved, and the stem cell filtering and separating efficiency is greatly improved.

The invention belongs to the technical field of cell separation, and relates to a method of quickly separating breast cancer primary tumor living cells in batches at a high purity. The method includes the steps of: (1) obtaining an isolated breast cancer fresh tissue sample; (2) performing pretreatment and quality control of the tissue sample; and (3) quickly separating the breast cancer primary tumor living cells in batches at a high purity, so that the high-purity breast cancer primary tumor living cells are separated in batches quickly. According to the invention, living breast cancer cells are separated from breast cancer tissue blocks with the purity of the tumor cells being higher than 90%. The separated breast cancer primary tumor living cells can be used in drug sensitive tests. The method overcomes the defects that the breast cancer primary tumor living cells are low in separation purity and are very liable to be polluted by fibroblast and can provide reliable research object for clinical drug sensitive tests. The invention provides the new method and wider material sources for database foundation of DNA, RNA and protein of primary tumor living cells and tumor in-vivo researching.

The invention relates to the technical field of single-cell isolation, in particular to a method for isolating single cells. With the method, size of single droplets can be accurately controlled by regulating size of laser energy, diameter of laser spots and thickness of photolytic material layers, only one cell is wrapped in each droplet, and isolation accuracy of the single cells is greatly improved; in addition, the laser energy does not act on the target cells direction, so that incompletion of cell functions cannot occur, and activity of the cells is sustained; meanwhile, the cells do not needed to be marked with fluorescent protein or magnetic particles, manual operation is omitted, and isolation efficiency is greatly improved.

The invention provides a centrifugal tube, and belongs to the technical field of cell separation. The centrifugal tube comprises an upper tube body, a lower tube body, a filtering membrane and a tubecap, wherein the upper tube body is of a structure with openings at the top end and the bottom end, the upper tube body is of a structure with an opening at the top end, the bottom end of the upper tube body is detachably inserted into the lower tube body, and the outer side wall of the upper tube body is in seal fit with the inner side wall of the lower tube body. The filtering membrane is plugged at the bottom end of the upper tube body and can allow separation liquid to pass and stop mononuclear cells from passing. The tube cap covers the top end of the upper tube body. As the filtering membrane can allow the separation liquid to pass and stop the mononuclear cells from passing, the separation liquid and red cells are positioned on the lower portion of the filtering membrane, an albuginea layer with blood plasma and the mononuclear cells are positioned on the filtering membrane and effectively isolated from the separation liquid and the red cells, and the purity of the separated mononuclear cells is ensured.

The invention provides a single cell separator, application thereof in a single cell separation process and a monoclonal cell preparation method, and relates to the technical field of single cell separation. The single cell separator comprises a liquid transfer needle and a liquid suction device, wherein the liquid transfer needle tightly communicates with the liquid suction device without leakage; and the liquid transfer needle comprises an intravenous injection needle which is bent at an angle of 75 to 90 degrees at a position 0.5 to 1 cm away from a needle opening. By using the single cellseparator provided by the invention, single cells can be quickly separated and extracted from cell sap, and the problems of complicated operation of a limited dilution method and high equipment requirement of a flow cytometer method in an existing single cell separation method are effectively relieved.

The invention provides a rapid and high-flux rabbit polyclonal antibody in-vitro production method,. According to the method, a B lymphocyte separation technology is mainly adopted to realize rapid, large-scale and stable production of rabbit polyclonal antibodies through in-vitro amplification culture, a B lymphocyte culture medium is optimized, tryptone is adopted, and through cooperation with blank rabbit serum, the B lymphocyte culture medium is prepared. The differentiation and proliferation amount of B lymphocytes are greatly improved, and a basis of cell quantity is provided for in-vitro production of the rabbit polyclonal antibodies. Through the method provided by the invention, the rabbits can be immediately subjected to blood sampling for production after being immunized for thethird time, the production cycle of products is greatly shortened, and the yield of the obtained rabbit polyclonal antibodies is 30 times or above of that of serum with the same volume in the traditional method. The method has the advantages of short cycle, high yield, low unit cost, stable quality and the like, and is suitable for large-scale production and application.