304 results about "CELL DEBRIS" patented technology

A method and apparatus for embedding cells that utilizes a flow-through embedding technique maximizes the efficiency of extractions and decreases time for embedding the cell fragments, minimizes cell loss, and automatically positions cell samples at the position in which a microtome blade will section them. The apparatus includes a cell flow pathway defined by an inflow tube for delivering cell fragments from a cell sample to a sample port. The sample port is in fluid communication with a tissue cassette having attached thereto a filter. The cell flow pathway is in communication with a reagent flow pathway for delivering the reagents through the sample port to the cassette. The apparatus is configured such that the application of pressure directs the cell fragments from the cell sample through the cell flow pathway, and effects delivery of the reagents through the reagent flow pathway. The apparatus produces an embedded cell block having concentrated cells near the plane of the block to be sectioned in a quick and efficient manner.

Methods for decellularizing mammalian tissue for use in transplantation and tissue engineering. The invention includes methods for simultaneous application of an ionic detergent and a nonionic detergent for a long time period, which may exceed five days. One method utilizes SDS as the ionic detergent and Triton-X 100 as the nonionic detergent. A long rinse step follows, which may also exceed five days in length. This long duration, simultaneous extraction with two detergents produced tissue showing stress-strain curves and DSC data similar to that of fresh, unprocessed tissue. The processed tissue is largely devoid of cells, has the underlying structure essentially intact, and also shows a significantly improved inflammatory response relative to fresh tissue, even without glutaraldehyde fixation. Significantly reduced in situ calcification has also been demonstrated relative to glutaraldehyde fixed tissue. Applicants believe the ionic and non-ionic detergents may act synergistically to bind protein to the ionic detergent and may remove an ionic detergent-protein complex from the tissue using the non-ionic detergent. The present methods find one exemplary use in decellularizing porcine heart valve leaflet and wall tissue for use in transplantation.

A substrate comprising a crosslinked polymer primer layer, and grafted thereto a ligand-functionalized polymer is provided. The grafted polymer has the requisite affinity for binding neutral or negatively charged biomaterials, such as cells, cell debris, bacteria, spores, viruses, nucleic acids, and proteins, at pH's near or below the pI's of the biomaterials.

A method and apparatus for a rapid disruption of cells or viruses using micro magnetic beads and a laser are provided. According to the method and apparatus for a rapid disruption of cells or viruses using micro magnetic beads and a laser, cell lysis within 40 seconds is possible, the apparatus can be miniaturized using a laser diode, a DNA purification step can be directly performed after a disruption of cells or viruses, and a solution containing DNA can be transferred to a subsequent step after cell debris and beads to which inhibitors of a subsequent reaction are attached are removed with an electromagnet. In addition, by means of the cell lysis chip, an evaporation problem is solved, vibrations can be efficiently transferred to cells through magnetic beads, a microfluidics problem on a rough surface is solved by hydrophobically treating the inner surface of the chip, and the cell lysis chip can be applied to LOC.

The invention relates to a modified spin column for the isolation and purification of plasmid DNA. A pre-filtration disc is included in a traditional spin column. During plasmid DNA isolation, the lysate can be loaded directly to the modified spin column, eliminates the need to first remove the flocculants containing cellular debris. This results in a much shortened process. Variation of the invention includes a depth filter in between the pre-filtration disc and the main separation matrix. Also provided are kits for isolation of plasmid DNA including the modified spin columns.

The present invention provides for decellularized tissue and method for decellularizing tissue. The method generally comprises the steps of obtaining a harvested tissue, performing a muscle shelf debridement, treating the tissue with an enzyme, washing the tissue with a detergent, and performing an organic solvent extraction on the tissue. The tissues decellularized according to the present invention have several advantages including removing more of the residual cell debris, dsDNA, and chemicals, as well as exhibiting less calcification and better ultimate tensile strength than tissues prepared not according to the method of the present invention.

Acellular amnion derived therapeutic compositions are described having a number of various compositional embodiments. An acellular amnion derived therapeutic composition has essentially no live or active amniotic stems cells. The amniotic stem cells may be destroyed, and the cells and cell debris may be removed from the acellular amnion derived therapeutic composition. An acellular amnion derived therapeutic composition may comprise micronized amniotic membrane particles, and / or amniotic fluid. An acellular amnion derived therapeutic composition may be a dispersion of micronized amniotic membrane combined with a fluid, such as plasma, saline, amniotic fluid, combinations thereof and the like. An acellular amnion derived therapeutic composition may be combined with a matrix component to form a composite. An acellular amnion derived therapeutic composition may be used in conjunction with a composition comprising viable cells, such as stem cells.

Guanidinyl ligand-functionalized polymers, methods of making the same, and substrates bearing a grafted coating of the ligand-functional polymers are described. The grafted polymer has the requisite affinity for binding neutral or negatively charged biomaterials, such as cells, cell debris, bacteria, spores, viruses, nucleic acids, endotoxins and proteins, at pH's near or below the pI's of the biomaterials.

The invention provides pipette tip extraction columns for the purification of a DNA vector from un-clarified cell lysate containing cell debris as well as methods for making and using such columns. The columns typically include a bed of extraction media positioned in the pipette tip column, above a bottom frit and with an optional top frit.

The invention discloses an extraction method of a specific mesenchymal stem cell exosome. The method comprises the following steps: collecting supernatant of serum-free mesenchymal stem cells: firstly, culturing mesenchymal stem cells, changing a culture medium for culturing, centrifuging, and removing cell debrises to obtain initial supernatant; taking the initial supernatant for carrying out further gradient centrifugation, transferring the centrifuged supernatant into a sterile centrifugal tube, centrifuging and collecting supernatant containing micro vesicles; centrifuging the supernatantcontaining the micro vesicles to obtain precipitate containing an exosome, washing and then centrifuging to obtain exosome precipitate; adding the obtained exosome precipitate into sterile PBS and filtering to obtain the specific mesenchymal stem cell exosome. The exosome collected in the extraction method disclosed by the invention adopts a filter membrane for filtration, so the extracted exosomeis enabled to be nanoscale; meanwhile, the rotational speed is reduced and optimized, high-speed mechanical damage is reduced, separation efficiency is high and the cost is low.

A method and apparatus for rapid disruption of cells or viruses using beads and a laser are provided. According to the method and apparatus for rapid disruption of cells or viruses using beads and a laser, cell lysis within 40 seconds is possible, the apparatus can be miniaturized using a laser diode, a DNA purification step can be directly performed after a disruption of cells or viruses, and a solution containing DNA can be transferred to a subsequent step after cell debris and beads to which inhibitors of a subsequent reaction are attached are removed with an electromagnet. In addition, by means of the cell lysis chip, an evaporation problem is solved, vibrations can be efficiently transferred to cells through magnetic beads, a microfluidics problem on a rough surface is solved by hydrophobically treating the inner surface of the chip, and the cell lysis chip can be applied to LOC.

A method for the indirect extraction of DNA greater than 300 kb in size from non-cultivatable organisms contained in an environmental sample is disclosed. The method comprises isolating the organisms from the sample and embedding the isolated organisms in a block of agarose where the organisms are subsequently lysed and the DNA subjected to a first alternating field electrophoresis to extract DNA fragments which are large in size and separate them from the cell debris. The first electrophoretic migration is followed by an enzymatic restriction step and additional electrophoretic migrations. The invention also encompasses DNA obtained by the method.

Acellular amnion derived therapeutic compositions are described having a number of various compositional embodiments. An acellular amnion derived therapeutic composition has essentially no live or active amniotic cells. The amniotic cells may be destroyed, and the cells and cell debris may be removed from the acellular amnion derived therapeutic composition. An acellular amnion derived therapeutic composition may comprise micronized placental tissue particles, and / or amniotic fluid. An acellular amnion derived therapeutic composition may be a dispersion of micronized amniotic membrane combined with a fluid, such as plasma, saline, amniotic fluid, combinations thereof and the like. An acellular amnion derived therapeutic composition may be combined with a matrix component to form a composite. An acellular amnion derived therapeutic composition may be used in conjunction with a composition comprising viable cells, such as stem cells.

A process for producing recombinant anti-botulinum toxin antibody comprising the steps of fermenting recombinant E. Coli cells in broth, concentrating the cells by removing the broth, crushing the concentrated cells, separating a permeate derived from the crushed cells from cell debris, purifying a recombinant antibotulinum antibody (Fab) from said permeate, and separating said Fab from impurities by diafiltration.

The invention provides a method for separating an exosome from a cell culture medium. The method comprises the following steps of: collecting fresh cell culture medium, centrifuging the collected cell culture medium to remove the suspended cell, other cell fragments and impurities in the culture medium, transferring the supernatant into a concentration tube for concentration, and adding Total Exosome Isolation (invitrogen) into the concentrated culture medium for the final extraction of the exosome. By adopting the method provided by the invention, the exosome in the cell culture medium can be conveniently obtained, and the separation cost is greatly reduced.

The invention discloses a Lactobacillus plantarum M1-UVs29 with an accession number of CGMCC No.2591. The Lactobacillus plantarum M1-UVs29 can be isolated from fermented meat products of various resources, such as preserved ham, sausage, salami, Xuanwei ham and the like, and is obtained after induced mutation and optimization. The Lactobacillus plantarum M1-UVs29 is a new strain of the induced mutation and isolation and has superior safety and minimal toxic and side effects, and can relieve classical clinical symptoms of coronary heart diseases, atherosclerosis, hyperlipemia and the like and treat cardiovascular diseases of various coronary heart diseases, atherosclerosis, hyperlipemia and the like incurred by hyperlipemia in a safe and effective way. The Lactobacillus plantarum M1-UVs29 is applied to the production of beverages, and health products of dairy products, fermented milk, acidsoy milk and the like and food additives. The obtained products of beverages, health products and food additives, containing the fermented Lactobacillus plantarum M1-UVs29 or metabolins, cell debris or secretions thereof, can effectively degrade cholesterol self-cumulated by human bodies and self-contained by food.

The invention discloses a method to produce bio-flocculant from residual activated sludge of a sewage treatment plant, pertaining to sludge treatment and disposition technique field in environmental engineering. The method is characterized in that: the residual activated sludge of the sewage treatment plant is used as material which is pre-treated for preparing purified sludge, and the purified sludge is cell-disrupted for preparing extraction serosity, which is solid-liquid separated to respectively obtain rough microorganism flocculant solution and cell debris residues. The rough microorganism flocculant solution is post-treated to prepare the concentrate solution of the microorganism flocculant or an finished product of a solid microorganism flocculant, and the cell debris residues is post-treated to realize the zero discharge of the residual activated sludge. The effects and benefits of the invention are that: sludge is regarded as a resource to be effectively employed; compared with other recycling way of the residual activated sludge, the invention has comparatively high added value of resource utilization. The generated bio-flocculant has good effects for removing the turbidity, the organic substances and the heavy metals of waste water.

A method for distinguishing erythroblasts from bacteria by automated hematology analyzers, such as, for example, the CELL-DYN® 4000 automated hematology analyzer and the CELL-DYN® Sapphire™ automated hematology analyzer. Bacterial cells scatter light and fluoresce differently than do red blood cells, white blood cells, erythroblast nuclei, and platelets. Signals generated by bacteria are distinguishable from those of erythroblasts because the signals generated by erythroblast nuclei are sufficiently unique that erythroblast nuclei can be distinguished from signals generated by bacteria. Signals generated by platelets, lysed red blood cell ghosts, and other cell debris are blocked by the triple-trigger circuitry of the hematology analyzer, because all of the signals generated by noise are below the AND / OR thresholds. Algorithm(s) in the software of the system detect and count signals generated by bacteria by means of the location and the shape of the signals generated by bacteria and calculate the concentration of bacteria per unit of body fluid. In addition, certain body fluids, such as, for example, synovial fluid, can be pretreated with a viscosity reducing agent for a short period of time to reduce the viscosity of the body fluid prior to analyzing a sample of the body fluid by an automated hematology analyzer.

The invention provides a method for preparing a mesenchymal stem cell exosome. The method comprises the steps that human umbilical cord tissue surface blood is cleaned to remove epidermal and vasculartissues, Wharton jelly is taken out for cleaning, then the tissues are cut into pieces, and inoculation is performed for culture; subculturing is performed; second generation of umbilical mesenchymalstem cells are inoculated and cultured; a cell stimulator factor for promoting secretion of the exosome is added; when the cells grow to the fusion degrees 90%, first cell supernate is sucked out forseparating the exosome; the first cell supernate is centrifuged to remove cell debris, and second cell supernate is collected; the first time of ultrafiltration and concentration is conducted on thesecond cell supernate, excessive moisture is removed, and a first concentrated solution is obtained; the first concentrated solution is subjected to the second time of ultrafiltration and concentration, excessive moisture is removed, and a second concentrated solution is obtained; the first time of ultracentrifugation is conducted on the second concentrated solution, and the supernate is taken andsubjected to the second time of ultracentrifugation, the supernate is removed, and precipitation is performed to obtain the exosome for stem cell secretion; normal saline is added for cleaning and centrifugation, the supernate is removed, and the exosome precipitate is obtained.

The invention provides a method for separating exosomes from animal plasma and for detecting purity. The method mainly comprises the following steps: first, thinning the plasma by virtue of a PBS (phosphate buffer solution) at the ratio of 1 to 1; removing cell components and cell debris in the plasma through centrifuging; precipitating the exosomes through ultrahigh-speed centrifugation; suspending the exosomes by virtue of 250[mu]l of the PBS solution; extracting total RNA in an exosome suspension by virtue of Trizol LS; and conducting poly (A) reversing and related gene quantification on the extracted total RNA, and detecting the purity. The method disclosed by the invention is conducive to researches on plasma exosome contents and exosome related functions in the future.

The invention discloses a method for extracting lipid from Chlorella sorokiniana CS-01 (CCTCC M209220), namely an ultrasonic-assisted chloroform / methanol extraction method. The method mainly comprises the following steps of: collecting algae, drying, extracting by using chloroform / methanol, performing ultrasonic crushing, washing, and drying. Through the certification, compared with a Soxhlet extraction method, an acid heat extraction method, and a chloroform / methanol extraction method, the method has the advantages that: the extraction ratio of the lipid is improved by 1.4, 1.7 and 0.5 timesrespectively; meanwhile, a lipid extraction phase is positioned under a cell debris solid phase, so the method is favorable for the subsequent direct separation of the lipid, and meets the requirements of mass industrialized production.

The invention provides pipette tip extraction columns for the purification of a DNA vector from un-clarified cell lysate containing cell debris as well as methods for making and using such columns. The columns typically include a bed of extraction media positioned in the pipette tip column, above a bottom frit and with an optional top frit.

In various aspects, the present invention provides novel and effective methods and kits for the isolation of sperm and sperm DNA from samples having at least one other cell type, or having the DNA of one other cell type. More specifically, a process for isolating sperm DNA from a mixture of sperm and non-sperm cells by filtration is provided. DNA from non-sperm cells is solubilized by selective lysis, and the intact sperm are retained on a filter, washed, and then treated in situ with a reducing agent to solubilize the sperm DNA. Significantly, centrifugation steps are not required, the process can be easily automated, and can be performed on many samples in parallel. The novel methods and kits are based on filtering selectively solubilized samples through filters that retain sperm by virtue of having uniform pore diameters that: are smaller than sperm, large enough to allow passage of cell debris and solubilized DNA; and are stable to pressure. The inventive methods and kits have broad utility, particularly in the forensic art.

The present invention relates to a method for isolating microorganisms and / or microorganisms nucleic acids from a bodily fluid that may comprise or may be suspected to comprise microorganisms and / or host cells and / or host cells debris. Microorganisms nucleic acids may further be isolated by lysing the isolated microorganisms. The present invention also relates to a method for detecting microorganisms in a bodily fluid. The present invention further relates to a saponin formulation and its use.

The invention relates to a preparation method of a mesenchymal stem cell exosome. The preparation method comprises the following steps: culturing mesenchymal stem cells for several hours by using a serum-free conditioned culture medium, centrifuging and removing the cells, collecting a liquid supernatant crude product, then re-centrifuging, filtering, and removing the residual cells as well as cell debris to prepare liquid supernatant containing an exosome; preparing the exosome by stepwise centrifugation of polyethylene glycol adopted by the liquid supernatant containing the exosomes. Exosome-expressed specific proteins CD63 prepared by the preparation method provided by the invention can effectively improve the mitochondria damage of the myocardial cells, prevent the cell apoptosis and improve the functions of the myocardial cells and are used for preparing a drug for treating myocardial ischemia reperfusion injury.

The invention relates to a spirulina phatensis polysaccharide and an extraction method thereof. The extraction method comprises the following steps: after multigelation and wall breaking of a spirulina phatensis powder suspension, centrifuging to obtain a cell lysate 1 and cell debris; adding ammonium sulfate in the cell lysate 1 until the saturation is 50%, after salting-out precipitation, centrifuging to remove phycobiliprotein to obtain a supernatant, and evenly mixing the supernatant and the cell debris to obtain a cell lysate 2; obtaining a spirulina phatensis polysaccharide crude extract by using a hot water extraction method; adding the crude extract to an ethanol / ammonium sulfate aqueous two-phase system, and extracting to obtain a bottom-phase solution of spirulina phatensis polysaccharide with higher purity; after the bottom-phase solution is desalted by dialysis, eluting by using a Sephadex G-150 chromatographic column and a DEAE Sephadex A-50 anion exchange column to obtain a pure spirulina phatensis polysaccharide solution; and freeze-drying, thereby obtaining a pure spirulina phatensis polysaccharide finished product. The extraction method can be used for avoiding the traditional complicated protein removal operation steps, has low cost, high yield, and high purity and activity of polysaccharide, and is suitable for intermittent and large-scale production and processing.

The invention discloses a novel serum-free medium. The serum-free medium comprises, by volume, 0.05-0.2 part of beta-mercaptoethanol, 0.5-2 parts of a non-essential amino acid aqueous solution, 4-6 parts of human mesenchymal stem cell cultural concentrated supernatant and 90-95 parts of a-MEM / DMEM-F12 and recombination human basic fibroblast growth factors with the final concentration of 5-15 ng / ml. A preparing method of the cultural concentrated supernatant includes the following steps that umbilical cord mesenchymal stem cell cultural supernatant is collected; cells, cell debris, impurities and the like are centrifugally removed; filtering is carried out through a micro-filtration membrane; ultrafiltration and concentration are carried out. When cultured through the medium, stem cells still keep the multi-directional differentiation potential and the high multiplication capacity under the long-time culturing condition.

Devices for expanded bed chromatography use inlet and outlet ports beneath a mesh that supports sorbent beads in a column. Placement of both ports beneath the mesh provides a horizontal “sweep flow” tangential to the mesh, to suppress the formation of particulate cakes on the lower surface of the mesh when liquids containing high particulate loads (such as cells or cell debris) are being processed. This design can be used with vibrators, hammering devices, intermittent reverse flow, or other means for disrupting the formation of particulate cakes or aggregates. Disrupters can also be used during elution, to accelerate the release of the valuable molecules from the sorbent. Initial tests indicate that these systems can efficiently handle heavily loaded liquids that would rapidly clog other systems previously known in the art.

The invention discloses a tumor-targeted delivery carrier based on cell-derived micro-vacuoles, a preparation method and an application. The preparation method comprises the following steps of: (A) preparing a conditioned medium: supplementing fetal bovine serum, antibiotics, DSPE-PEG-Biotin and DSPE-PEG-Folate into a basal medium; (B) using the obtained conditioned medium in cell culture, and collecting cell culture supernatant for subsequent separation; (C) carrying out low-speed centrifugation on the obtained culture supernatant to remove cell debris and apoptotic bodies, then adding SA-IONPs, mixing uniformly, incubating, then separating by a magnet, using PBS for re-suspension, eluting for multiple times to obtain the cell-derived micro-vacuoles with membrane surfaces modified by folic acid and iron oxide nano-particles, and freezing for storage; and (D) loading chemotherapeutic drugs or therapeutic genes into the functionalized micro-vacuoles doubly-modified by an electroporation mode, and carrying out re-suspension after separation with the magnet. The tumor-targeted delivery carrier based on cell-derived micro-vacuoles, the preparation method and the application disclosed by the invention are applicable to specific targeting delivery of multiple chemotherapeutic drugs and therapeutic genes, and have the advantages of enhancing the anti-tumor effect, reducing the systemic toxicity and improving the clinical effect of the current therapeutic selection, so that a new hope is brought for clinical therapy of tumors.