21 results about "Ficoll" patented technology

The invention relates to the field of blood detection reagents, in particular to a blood free DNA stabilizer and application thereof. Every 100 ml of the blood free DNA stabilizer comprises 0.5-5 g of a buffer agent, 2-20 g of an aldehyde fixing agent, 15-20 g of an anticoagulant, 10-50 g of a cell stabilizer, 2.5-12.5 g of a nuclease inhibitor, and the balance of ultrapure water. The pH value of the blood free DNA stabilizer is 6.8 to 7.8; the cell stabilizer is selected from one or two or more than two of polysucrose-50, polysucrose-70, polysucrose-400, polyethylene glycol-4000, polyethylene glycol-6000, polyethylene glycol-8000 and glycine betaine. Preferably, the cell stabilizer is selected from polysucrose-70, and every 100 ml of the blood free DNA stabilizer contains 10 g of polysucrose-70. The stabilizer provided by the invention is low in toxicity and low in cost, and can maintain free DNA stability and inhibit blood cell genome release at normal temperature for 10 days, so that the storage cost and the transportation cost of a blood sample are effectively reduced, and the detection accuracy of the blood free DNA is improved.

The invention provides a method for preparing autoserum antigen-sensitized DC-CIK cells, which comprises the following steps: (1) preparation of autoserum: collecting peripheral blood of a patient, standing and centrifuging, and collecting supernatant to obtain the autoserum; (2) separating the PBMCs: separating the PBMCs from the peripheral blood of the patient by using a Ficoll-Paque separating medium to obtain the PBMCs; (3) preparing DC cells: culturing PBMCs, adding GM-CSF and IL-4 to induce DC differentiation, and then adding TNF-alpha and an autoserum antigen to promote DC maturation; (4) preparation of CIK cells: culturing PBMCs through IFN-gamma, an anti-CD3 monoclonal antibody and IL-2, and inducing generation of the CIK cells; and (5) co-culture of the DC-CIK cells: co-culturing the mature DC cells and the CIK cells in proportion, and adding GM-CSF, IL-2 and anti-CD3 monoclonal antibodies to promote cell proliferation and activation so as to obtain the autoserum antigen sensitized DC-CIK cells. The autoserum antigen sensitized DC-CIK cell provided by the invention has higher specificity and safety, and the effect of tumor immunotherapy is remarkably improved.

The invention discloses a primary culture method of mesenteric interstitial cells, which relates to the technical field of cell culture, and is characterized by comprising the following steps: selecting mice with a growth cycle of more than 4 weeks, separating mesenteric tissues of the mice, treating the mesenteric tissues by using mesothelial cell dissociation liquid to remove mesothelial cells; the method comprises the following steps: separating mesenteric interstitial cells by using a physical cutting and enzyme digestion method, purifying the cells by using a Ficoll / Percoll density gradient centrifugation method, finally carrying out multiplication culture on the mesenteric interstitial cells in a culture medium containing itaconic acid derivatives, and carrying out passage operation when the cell confluence degree reaches 80%. The cell culture method provided by the invention effectively avoids mixing of mesothelial cells, endothelial cells and other cells, can promote the mesenteric interstitial cells to adhere to the wall and grow as soon as possible, improves the cell amplification rate, and reduces the cell culture cycle.

The invention aims to provide a method for separating and amplifying placenta decidua matrix cells in a perinatal period. The method comprises the following steps: adding decidua tissues into a culture medium A containing collagenase for digestion treatment; cleaning to obtain a cell suspension A, and sieving to obtain a cell suspension B; mixing the cell suspension B with a Ficoll solution, and carrying out primary gradient centrifugation; mixing the cell layer and the supernate layer with normal saline, and then carrying out secondary gradient centrifugation; and culturing the cell block mass and the culture medium B in an incubator. According to the method, the success rate of separation and amplification of the placenta decidua interstitial cells in the perinatal period can be increased, the separated and proliferated cells have group cell biomarker expression for tissue repair, and the separation technical method is improved, so that highly purified cells are obtained.

Provided is an aqueous solution for separating blood cells, the aqueous solution being capable of suppressing a change in properties even when sterilized by radiation.　This aqueous solution for separating blood cells comprises: polysucrose (A); diatrizoic acid and / or a metal salt thereof (B); and carboxylic acid different from diatrizoic acid and / or a metal salt thereof (C).

The invention discloses an ovarian tissue low-temperature cryopreservation solution as well as a preparation method and application thereof, and relates to the technical field of biomedical materials. The invention relates to an ovarian tissue low-temperature cryopreservation solution as well as a preparation method and application thereof. Every 100 ml of the ovarian tissue low-temperature cryopreservation solution comprises 15-25 ml of dimethyl sulfoxide, 15-25 ml of ethylene glycol, 15-20 g of polysucrose, 38-46 g of sucrose, 40-45 ml of a phosphate buffer solution and 0.12-0.13 g of human serum albumin. According to the low-temperature cryopreservation solution for the ovarian tissues, the dimethyl sulfoxide and the ethylene glycol are used as permeable low-temperature protective agents to prevent the ovarian tissues from being damaged at low temperature, and the polysucrose and the sucrose are used for regulating the cell osmotic pressure to improve the cell dehydration effect, reduce the formation of ice crystals in cells and increase the viscosity of the solution; according to the low-temperature cryopreservation solution for the ovarian tissue, the solution forms an amorphous glassy state instead of ice crystals in the freezing process, the human serum albumin maintains osmotic pressure and nutrition, the tissue survival rate and the functional integrity are guaranteed, the ovarian tissue can be efficiently cryopreserved through the low-temperature cryopreservation solution for the ovarian tissue, and the follicle survival rate and the functional integrity after ovarian transplantation are effectively improved.

The application discloses a double-frequency ultrasonic-assisted extraction method of fig polysaccharide and application, and belongs to the technical field of natural product extraction. The method comprises the following steps: S1, preparing an ethanol solution of fig powder; S2, placing the solution in a double-frequency ultrasonic device for ultrasonic-assisted extraction, and the ultrasonic conditions comprise the following: an ultrasonic frequency of 40 kHz and 25 kHz, an ultrasonic power of 345-525 W, and an ultrasonic time of 4-12 min; and S3, obtaining fig polysaccharide extract after filtration. The method is an efficient, rapid and simple fig polysaccharide extraction method, and is particularly suitable for an industrialized production scene with strict requirements on extraction time, and the extracted fig polysaccharide has good in-vitro antioxidant activity, can be used for preparing antioxidant-related functional food, medicine and cosmetics, and has a good development prospect in the field of health products.

The invention discloses a method for separating and extracting PBMC (peripheral blood mononuclear cells), which comprises the following steps: on the premise of retaining PBMC in a sample, carrying out volume reduction treatment on a sample solution, namely, carrying out volume reduction on the sample solution to discharge a certain amount of plasma and red blood cells, and then carrying out ficoll solution centrifugation, so as to carry out one-time PBMC separation operation on a large-volume sample; the process flow time of PBMC separation and extraction is greatly shortened, the cell yield is high, the activity is high, and an effective cell acquisition way is provided for clinical and basic cytology research so as to facilitate subsequent immune cell experiments; according to the separation and extraction method, less separation liquid, washing liquid (replacement liquid) and disposable consumables are used, so that the time, consumables and reagent cost are saved.

The invention discloses a DC-CIK co-culture method which comprises the following steps: S1, collecting blood of a tumor patient, and separating PBMC cells in the blood of the tumor patient by adopting a Ficoll separation method; s2, resuspending the separated cells by using a culture solution, then transferring the cells into a T75 bottle, and culturing the T75 bottle in a carbon dioxide incubator; s3, culturing for 2.5-3.5 hours in a carbon dioxide incubator, pouring supernatant into a new T175 bottle, adding a CIK factor 1 to culture CIK cells, and adding a DC starting factor into a T75 bottle to culture DC cells; s4, continuing to culture for 20-26 hours, and adding a CIK factor 2 into the T175 bottle; s5, continuing to culture for 110-130 hours, and adding a DC maturation factor into the T75 bottle; s6, continuing to culture for 160-170 hours, harvesting the matured DC cells, and transferring the DC cells and the CIK cells into a culture bag for culture; and S7, culturing for 330-340 hours, harvesting, and testing the DCCIK flow phenotype. The DC-CIK cell solves the problems of low purity, poor cell killing function and long tumor killing period of the existing DC-CIK cell.

The invention discloses a method for rapidly separating and culturing preadipocytes in carp muscle and inducing adipogenesis, and relates to a method for rapidly separating and culturing fish cells and inducing adipogenesis. The isolated culture comprises the following steps: 1, muscular tissue acquisition; 2, digesting muscular tissues; 3, purifying the preadipocytes by Ficoll; 4, counting the cell suspension; and 5, purifying by adopting a differential adhesion method. And fat formation induction: performing induction by using a fat formation induction culture medium, and successfully performing fat formation induction on the proadipocytes in the carp muscle. The invention provides a method for jointly digesting muscular tissues through combination of pancreatin and collagenase, and fish intramuscular preadipocytes with high purity and large quantity are effectively obtained. The carp intramuscular preadipocytes obtained by the invention are rapid and efficient in adipogenesis, and a basic material is provided for constructing a carp intramuscular fatty degeneration model.

The present invention relates to a separating solution for separating shark PBMC, which contains 1-2 g of NaCl in 100 mL of Ficoll-Urografin layering solution, and the density of the separating solution is 1.087 g / mL - 1.097 g / mL. The separation method is to mix fresh anticoagulated whole shark blood with shark blood diluent, then combine it with the separating solution, and after centrifugation, a PBMC layer with obvious stratification is obtained. After multiple cell resuspensions and centrifugal separations, high-purity shark PBMC is obtained. The separating solution has simple components, the separation operation is simple and easy to perform, and each component is a very stable substance. The prepared separating solution is easy to store, which simplifies the preparation process and storage conditions.

The invention provides a preparation method of nano-particles capable of regulating the crowding degree in cells, which comprises the following steps: firstly preparing degradable silicon dioxide nano-particles BSN (at) Ficoll containing disulfide bonds entrapped with polysucrose, then modifying amino at the tail end to obtain BSN (at) Ficol-NH2 nano-particles, and then modifying with folic acid and hyaluronic acid to obtain FA-HA-BSN (at) Ficoll nano-particles, namely FHS (at) Ficoll for short. According to the FHS-Ficoll nano-particle disclosed by the invention, the polysucrose is used as a crowding agent, the silicon dioxide is used as a core skeleton, the disulfide bond is used as a polysucrose responsive release switch, and the folic acid and the hyaluronic acid are jointly used as targeting ligands, so that accurate targeted delivery of proinflammatory macrophages and glutathione responsive release crowding agents can be realized, and the crowding degree in cells can be dynamically regulated; and protein is induced to generate phase separation, so that the relieving of inflammatory response can be effectively promoted, and the repair and regeneration process of damaged tissues is accelerated.

The present invention discloses a method for establishing a pancreatic cancer screening model based on mass spectrometry flow cytometry, relating to the medical field. The method comprises the following steps: treating a peripheral blood sample with Ficoll separation to obtain PBMCs, and performing mass spectrometry flow cytometry analysis on the PBMCs to obtain a flow cytometric analysis data set; processing the flow cytometric analysis data set using a random forest algorithm, and performing feature screening using the negative proportion of cell surface markers and subpopulation proportions as modeling features to obtain a pancreatic cancer screening model. The present invention can be used for screening and auxiliary diagnosis of pancreatic cancer.

The invention relates to a Taq enzyme antibody blocking stabilizer, which is prepared from 0.01 to 1 percent of N, N-dihydroxyglycine, 0.01 to 0.5 percent of polysucrose 70, 0.01 to 1 percent of glucan and 0.01 to 2 percent of polyvinylpyrrolidone, and the pH (Potential of Hydrogen) of the Taq enzyme antibody blocking stabilizer is 7 to 9. The Taq enzyme antibody blocking stabilizer provided by the invention solves the problems of non-specificity and performance reduction in a thermal acceleration process and / or a long-term storage process of a fully premixed reaction system of high-sensitivity qPCR containing primers, so that the debugging difficulty of the fully premixed reaction system is greatly reduced. The Taq enzyme antibody blocking stabilizer disclosed by the invention is suitable for different Taq enzymes.

The invention discloses a method for enhancing anti-tumor activity of NK cells, and relates to the technical field of anti-tumor treatment of NK cells, peripheral blood mononuclear cells are separated through a Ficoll density gradient centrifugation method, and CD56 + CD3-NK cells are purified after IL-2 amplification. Then, the NK cells are co-cultured with IL-21 (interleukin-21), DZNp (2, 3-phthalazinone) and resveratrol to promote immune activation, epigenetic regulation and antioxidation of the NK cells, so that the anti-tumor capability of the NK cells is remarkably enhanced. Through the synergistic effect of the IL-21, the DZNep and the resveratrol, the anti-tumor effect of the NK cells is maximized. IL-21 activates immune response, DZNep enhances cytotoxicity, resveratrol alleviates oxidation pressure, and IL-21, DZNep and resveratrol synergistically optimize immunocompetence and anti-tumor ability of NK cells and overcome a tumor immune escape mechanism.

The invention discloses a method for separating mononuclear cells from alpaca peripheral blood, a preparation and a preparation method, and relates to the technical field of biology, the preparation comprises a reagent A and a reagent B, the reagent A comprises 1.0 mg / mL to 4.0 mg / mL of hydroxypropyl methylcellulose, 2.0 mg / mL to 6.0 mg / mL of chitosan quaternary ammonium salt, 48.0 mg / mL to 54.0 mg / mL of polysucrose 400 and 80.0 mg / mL to 10.0 mg / mL of meglumine diatrizoate; the reagent B comprises a Percoll separating medium, a diluent of the Percoll separating medium is normal saline or PBS, and the density of the Percoll separating medium is 1.079 g / mL-1. 081 g / mL. The problems that in the prior art, the density of neutrophils and the density of mononuclear cells in alpaca peripheral blood are close, the mononuclear cells are separated and extracted by using a single or discontinuous gradient separating medium, and the separation efficiency of the neutrophils and the mononuclear cells is low can be solved. And the effect is poor.

The utility model relates to the technical field of cell separation, and discloses a centrifugal tube for quickly separating peripheral blood bone marrow mononuclear cells, which comprises a centrifugal tube, an upper internal thread is arranged on the inner wall of the centrifugal tube, and a lower internal thread is arranged on the inner wall of the centrifugal tube. Blood and ficoll liquid are isolated through the filter membrane sheet and are prevented from being mixed too much to affect the separation effect, pure mononuclear cells can be simply, conveniently and rapidly collected in the centrifugal process due to the isolation effect of the inert separation gel and the blocking function of the filter membrane sheet, and after centrifugation is completed, the mononuclear cells are collected in a direct and rapid pouring mode, so that the separation effect is greatly improved. The inert separation gel is used for isolating centrifuged red blood cells, preventing the red blood cells from being mixed into separated mononuclear cells when supernatant is poured out, improving the separation method of the mononuclear cells, improving the separation efficiency of the mononuclear cells, reducing the experiment difficulty, saving the experiment time and guaranteeing the reliability of experiment results.

The invention relates to a use method and application of a mitochondrial membrane potential probe. The use method comprises the following steps: (a) separating human peripheral blood mononuclear cells by adopting a Ficoll method; (b) carrying out flow antibody staining; (c) centrifugally collecting the cells, cleaning with a PBS buffer solution, and adding a cell culture medium to resuspend the cells; (d) adding a fluorescent probe JC-1 into the culture medium, incubating in a 5% CO2 cell incubator at 37 DEG C for 15-30 minutes, centrifuging, precipitating cells, and discarding the supernatant; adding resuspended cells, centrifuging, precipitating the cells, and discarding the supernatant; and (e) resuspending the cells in a precooled PBS buffer solution, and measuring and analyzing the membrane potential in a flow cytometer. The progress of the liver disease can be analyzed according to the significant difference, the accuracy and specificity of diagnosis are significantly improved, and a reliable basis is provided for clinical diagnosis.

This invention provides an aqueous solution for blood cell separation that can suppress changes in properties even after sterilization by radiation. The aqueous solution for blood cell separation according to the present invention comprises polysucrose (A), diatrizoic acid and / or its metal salt (B), and a carboxylic acid and / or its metal salt different from diatrizoic acid (C).

The invention relates to the technical field of molecular diagnosis, in particular to a multiple reverse transcription polymerase chain reaction detection reagent buffer solution and a kit. According to the invention, multi-factor comprehensive research is carried out on multiple reverse transcription polymerase chain reactions, especially on the performance of a one-step RT-PCR reaction system constructed by taking Z05 enzyme as a bifunctional enzyme raw material, so that the influence of a polyhydroxy compound as an additive on various performance indexes of the reaction system is fully analyzed; a specific polyhydroxy compound (Ficoll 400) is creatively selected to be compounded with pinotriose to serve as a reaction buffer reagent additive, a synergistic interaction effect is generated, and especially under a buffer reagent system preferably constructed by a Tricine buffer solution, the final fluorescence value of an RT-PCR fluorescent quantitative reaction system is more efficiently increased, the signal-to-noise ratio is increased, and the detection accuracy is improved.

The invention provides a mass spectrum flow cytometry detection kit for evaluating the immune state of a patient with non-Hodgkin lymphoma and application, and the mass spectrum flow cytometry detection kit covers 36 antigens and can cover at least more than 90 immune cell subgroups. Comprising the following steps: treating a peripheral blood sample by using a Ficoll separation method to obtain PBMC (peripheral blood mononuclear cells), and performing mass spectrometry flow analysis on the PBMC to obtain a flow analysis data set; a flow analysis data set is processed through a random forest algorithm, the immune cells in the peripheral blood of the NHL patient are rapidly and accurately subjected to deep analysis, and the relation between the peripheral blood immune cell composition and function and immunotherapy scheme selection and curative effect is explored. According to the invention, through multi-dimensional cell subset analysis, treatment scheme selection and curative effect evaluation suggestions are provided for lymphoma immunotherapy patients.