99 results about "Cell yield" patented technology

Methods for the maximizing parameter of the in vitro growth and expansion of mammalian cells, specifically postpartum-derived cells in containers such as roller bottles is described. Methods of optimizing growth rate and cell yield in such culture systems are provided. The methods are particularly adapted for human postpartum-derived cells, such as umbilicus-derived cells.

The invention relates to a plate-type photobioreactor, which comprises at least one flow channel, wherein each flow channel comprises at least 2 upper baffle plates, which are arranged on the inner wall face on a light receiving side, and at least 2 lower baffle plates, which are arranged on the inner wall face on a backlight side; and the included angles of the length directions of the upper and lower baffle plates and the flowing direction of culture solution are 20 to 70 DEG and are opposite. The plate-type photobioreactor has a specific internal structure, allow microalgae cells to move back and froth between the bright and dark areas in the photobioreactor, and give play to the 'flash effect' of the microalgae cells, thereby improving the light energy utilization and microalgae cells yield of the microalgae large-scale culture. The plate-type photobioreactor disclosed by the invention is suitable for large-scale culture of various microalgae cells.

The invention relates to the preparation and application method of a medium, which is specifically related to a medium which is used for Vero cell and a culture method of the Vero cell. The invention belongs to the biological product area. The culture method of the invention is that the adhering Vero cells are suspend and cultured in SFM serum-free medium for the domestication and adaptation; the growing pattern of the Vero cell changes from adherence growth to suspension growth; the Vero cells are further amplified in Vero amplification CHEMICALLY DEFINED MEDIUM. The method of the invention has good effects; the growing pattern of the Vero cell is transformed from adherence growth to suspension growth, which changes the growth pattern of the cell and greatly increases the yield of the cell.

The invention relates to a culture method for efficiently obtaining adipose mesenchymal stem cells, and solves the problem that the cell viability obtained in the prior art is low. The culture method comprises the following steps: separation of the adipose mesenchymal stem cells: washing D.Hanks with fat tissues, detecting to guarantee no pollution, adding a solution mixture of I-type collagenase and trypLETM digestive enzyme with several times of volume, and stopping enzymolysis after carrying out rotary vibration digestion for dozens of minutes at the temperature of 37 DEG C, centrifuging to remove a liquid supernatant, and filtering with screen cloth to obtain an adipose mesenchymal stem cell suspension; culture of the adipose mesenchymal stem cells: inoculating a culture bottle bloodless culture medium with the adipose mesenchymal stem cell suspension, carrying out primary cell culture, observing with an inverted phase contrast microscope in the culture period, and changing a fresh culture medium, when the cells are cultured to 80% to 90% fusion, extensively inoculating a new culture bottle with the cells for culture after digestion with trypLETM, and subculturing for several generations to obtain the purified mesenchymal stem cells. The culture method has the advantages that the digestion is more sufficient, the cell yield is higher, the mesenchymal stem cells are protected from being damaged, and the viability is high.

This invention relates to embodiment to a molecular beacon, methods and kits for the detection of expression of p21(Waf1 / Cip1), in response to chemotherapy. Specifically, the introduction of a phosphorothioate-modified p21-beacon by transfection in human tumor cells yields increased signal in a dose dependent manner.

The present invention discloses a method of isolation, pooling and further culturing of Mesenchymal Stem cells (MSC) for clinical application. Present invention also discloses the method of establishing Master Cell bank, followed by Working Cell Bank from which the final therapeutic composition referred to as Investigational Product / Investigational Medicinal Product comprising of allogenic bone marrow-derived MSC is formulated for clinical applications. Present disclosure also discloses a robust manufacturing process for consistent production of clinical grade Mesenchymal Stromal cells (MSCs). The process enables production of highly viable potent cells. The process steps relating to preparation of media, cell seeding, harvesting are fine tuned to achieve consistency in cell yield, superior cell viability, purity, improved cell proliferation, high cell recovery, low HLA-DR expression, reduction in culture duration. The viability and purity of cells are further improved by optimized wash process without cell loss / cell stress. The disclosure further provides a method of cyrostoring MSCs at high cell density without affecting the viability of cells. It further provides economical means to store and transport at −80° C.

The present invention relates to a method of increasing the yield of cells collected in a urine sample comprising the steps of applying energy wave forms to a bladder wherein the energy loosens cells attached to the inner lining of the bladder, and collecting urine sample containing the loosened cells.

The invention discloses a method for separation and in-vitro culture of pancreatic cancer tissue organs of human. The method comprises the following steps that (1) after the pancreatic cancer tissue of human is cut, enzymic digestion is repeatedly carried out for 10-20 minutes at 37 DEG C in a rotation speed of 30-40 rpm three times, and a pancreatic cancer single cell is obtained; (2) the pancreatic cancer single cell and matrigel are uniformly mixed, then a culture medium containing a growth factor PEG2 and gastrin I are added for culture, and the pancreatic cancer tissue organs of human areobtained. The method for separation and in-vitro culture of the pancreatic cancer tissue organs of human has the advantages that the tumor tissue is digested by adopting a short-time repeated digestion method, which can effectively improve the cell yield and cell viability; the culture medium containing the growth factor PEG2 and the gastrin I is adopted for culture, which can effectively promotethe growth of the pancreatic cancer organs and increase the survival rate of the pancreatic cancer organs.

The invention provides methods for differentiating pluripotent stem cells such as ES cells with improved progenitor and differentiated cell yield using low oxygen conditions and optionally in the absence of exogenously added differentiation factors.

The invention provides a method for producing viral vaccines, which comprises the following steps of: 1) inoculating cells for producing virus into a bioreactor with a Disks carrier, and culturing; and 2) harvesting the virus to prepare the vaccines. In addition, besides the step 1) of culturing the cells for producing the virus, the method further comprises the following steps of: when the cellsfor producing the virus grow to a certain density, inoculating and proliferating the virus to be produced. By the method, the cell yield can be improved, the production cost can be reduced, and the production efficiency can be improved.

The application discloses a preparation method of a fish single-cell suspension. The preparation method comprises the following steps: preparing a PBS reagent, a D-Han liquid and an IV type collagenase liquid, preparing a kidney tissue block of a fish, and washing the kidney tissue for more than three times with PBS; putting the kidney tissue block in a flat vessel of an ice bath, adding PBS, shearing the kidney tissue block into small pieces, and putting the tissue blocks in a high throughput tissue centrifuge tube of a burnisher; adding the IV type collagenase liquid into the centrifuge tube and meanwhile, adding PBS to fix the volume to 4ml, putting a zirconium oxide wall to grind, and performing enzymatic digestion on a tissue homogenate; and performing filtration, centrifugalization, washing with PBS, performing centrifugalization to abandon supernate and removing cell debris, filtering the cell suspension and fixing the volume to 3ml with PBS, and storing the cell suspension for later use at 4 DEG C. By adopting an enzymatic hydrolysis grinding method, advantages of a griding method and an enzymatic hydrolysis method are combined. The tissue is more fully treated by enzymatic hydrolysis grinding, so that the problem that the cell yield is low and the cell viability is low for preparing the fish single-cell suspension can be solved.

The invention relates to the field of stem cell culture and in particular relates to a separation method of limbal stem cells. The separation method has the beneficial effects that limbal tissues are subjected to enzymolysis under the combined action of a compound enzyme by combining neutral protease with collagenase IV, thus ensuring obtainment of the cells through enzymolysis in a short time, reducing damages to the cells, increasing the cell yield and improving the purity of the obtained cells; the method is simple and convenient to operate, dispenses with high-end instruments and equipment and causes smaller damages to the cells; experiments show that by adopting the method provided by the invention to separate the limbal stem cells, every 1cm<3> of limbal tissues can obtain 8.4*10<5>-9.0*10<5> limbal stem cells, the positive rate of an antibody CX43 in the cells is 0, the positive rate of AE5 is 0.1%, the positive rate of BrdU is 70.1%, and the positive rate of PCNA is 82.1%; the separation method has obviously better effects than the prior art.

The invention provides digestive enzyme liquid for poultry tibial growth plate cartilage primary cell separation. A complete medium is used as basic liquid, and is prepared from the following ingredients including collagenase IV with the mass concentration being 80 to 100mg / mL and hyaluronidase with the mass concentration being 80 to 100mg / mL; the complete medium takes a high-glucose DMEM-F12 culture medium as a basic culture medium, and contains the following ingredients including fetal calf serum with the volume concentration being 8 to 12 percent, vitamin C with the mass concentration being 4.5 to 5.5mg / mL and mycillin with the mass concentration being 1 to 10mg / mL. Through the selection on digestive enzyme and the concentration optimization, the cell yield reaches 85 to 95 percent or higher; during the further culture on the digested cells, the cell upward trend is good; meanwhile, the cell viability of the digested primary cell reaches 85 to 95 percent or higher; the cell viability is relatively stable.

The invention discloses an N-type high-efficiency heterojunction solar cell and a preparation method thereof. The N-type high-efficiency heterojunction solar cell is made of an N-type silicon wafer. The preparation method comprises the following steps of: firstly, performing gettering treatment on an N-type silicon wafer, and then completing conventional manufacturing of the heterojunction solar cell. The gettering treatment comprises the following steps: (1) cleaning a silicon wafer; (2) coating or depositing a layer of gettering source on the front surface and the back surface of the siliconwafer; (3) performing heat treatment on the silicon wafer coated or deposited with the gettering source through a chain annealing furnace to finish gettering, and forming a gettering layer on the surface of the silicon wafer; and (4) conducting corroding to remove the gettering layer on the surface of the silicon wafer. According to the invention, the content of metal impurities in the silicon wafer body is reduced, the minority carrier lifetime of the silicon wafer is prolonged, and the conversion efficiency of the cell is finally improved; the cell efficiency distribution is more centralized, and the product consistency is improved; and the cell edge electric leakage rate is reduced, and the cell yield is improved.

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 discloses a method for extracting adipose-derived stem cells. The method includes steps of A, pretreating adipose tissues; B, adding type-I collagenases into the pretreated adipose tissues and carrying out digestion; C, collecting digested adipose cells positioned on lower layers after digestion is carried out for preset time; D, aspirating the type-I collagenases and undigested adipose cells positioned on upper layers and carrying out digestion again; E, repeatedly executing the step C and the step D until pretreated adipose cells are completely digested; F, culturing a plurality of generations of digested adipose cells to obtain passage cells; G, collecting the passage cells and carrying out a plurality of items of cell detection; H, transferring passage culture cells intocell seed banks after the passage cells are qualified in the cell detection items, and storing the passage culture cells in the cell seed banks. The method has the advantages that influence of the digestion time on the adipose-derived stem cells can be effectively lowered in step-by-step digestion modes, and the cell yield or the content of the adipose-derived stem cells can be increased.

The invention relates to an isolated culture method of piglet myocardial fibroblasts and belongs to the technical field of cell culture in modern biotechnology. The method includes: 1, collecting cardiac tissues from a piglet 1 to 3 days old, shearing ventricular tissues, performing PBS (phosphate buffer saline) pre-cooling, and rinsing several times; 2, digesting the myocardial tissues, namely performing multiple digestion at 37 DEG C with a mixture of trypsin 0.25% and collagenase II 0.1% having a ratio of 1:1, adding DNA deoxyribonuclease (0.02 mg / mL) to reduce cell suspension viscosity and increase cell yield, and adding red blood cell lysis buffer to reduce the amount of red cells; 3, culturing piglet myocardial fibroblasts, namely re-suspending cells with DMEM (Dulbecco modified Eagle medium) high-glucose medium containing fetal calf serum 10-15%, and performing differential attachment to obtain the myocardial fibroblasts. The method is simple and easy to master, time efficient and high in success rate, the obtained piglet myocardial fibroblasts are good in form, good in activity and abundant. Certain basis is provided for establishing a cell model of piglet myocardial fibroblasts to study related diseases such as cardiac hypertrophy in future.

The invention provides a cryoprotectant capable of being directly transfused back for stem cells and related products thereof, and belongs to the field of cryopreservation protective agents. The cryoprotectant solves the problems that animal-derived raw materials are added into an existing cryoprotectant, uncertain factors are brought to the self-renewal and differentiation process of cultured cells, and the like, and the cryoprotectant used for stem cells and related products thereof and capable of being directly reinfused comprises the following components in parts by volume: 5-10 parts of dimethyl sulfoxide, 30-35 parts of a compound electrolyte injection, 30-35 parts of glucose injection, 15-20 parts of human serum albumin injection and 10-15 parts of dextran injection. The cryoprotectant has the advantages that the stem cells can be directly transfused back after being revived, cleaning is not needed, time and cost are saved, and the cell yield and cell viability can be improved.

The invention discloses a culture solution and a differentiation method for differentiating pluripotent stem cells into natural killer cells. The culture solution comprises a DMEM / F12 culture medium and a blood cell culture medium in a volume ratio of 1: (0.9-1.3), and further comprises an induction factor, L-ascorbic acid, L-glutamine, L-leucine, human serum albumin, transferrin, beta-mercaptoethanol, sodium selenite, ethanolamine, a trace element A and a trace element B. The culture solution provided by the invention can induce differentiation of the pluripotent stem cells towards the NK cells more efficiently, and the pluripotent stem cells can be directionally differentiated and cultured in a short time by using the method provided by the invention to obtain the NK cells. The NK cell differentiation method is free of animal-derived components, high in NK cell yield and simple in differentiation process, a differentiation period is shortened to 14 days, NK cell yield is greatly increased, the NK differentiation period is shortened, the NK differentiation process is simplified, and preparation cost is reduced.

The invention provides methods for differentiating pluripotent stem cells such as ES cells with improved progenitor and differentiated cell yield using low oxygen conditions and optionally in the absence of exogenously added differentiation factors.

The invention relates to a gradient illumination photobioreactor and application thereof. The photobioreactor has a box-type structure, and is divided into a compartments I, compartments II and compartments III, which have light intensity from high to low; a pair of compartment I, a pair of compartment II, and a pair of compartment III are arranged in each reactor box body; liquid flows through an overflow pipe or an overflow port between the compartments; microalgae is cultured in 3 compartments with high intensity from high to low, and the microalgae liquid concentration and aeration amount are increased correspondingly according to the light intensity from low to high; and the photobioreactor with gradient light and the cultivation of microalgae can improve the conversion rate of light energy; and for the continuous culture of microalgae, the photobioreactor is equivalent to multiple complete-mixing-flow-reactors in series, and can improve the cell yield.

The invention discloses a method for culturing primary gill cells of takifugu obscurus. The method comprises the following steps: (1) taking a gill tissue of the takifugu obscurus, soaking with a soaking solution containing double-resistant penicillin and streptomycin, cleaning with a cleaning solution, and cutting gill filaments; (2) digesting the gill filaments by using trypsin, filtering, centrifuging and taking precipitates; and then removing red blood cells by using a red blood cell lysate, and centrifuging to obtain cell precipitates; and (3) adding a complete culture solution into the cell precipitates, blowing the cells, supplementing the complete culture solution, and culturing. Compared with the prior art, the method provided by the invention is applied to culture of the primarygill cells of the takifugu obscurus, tissues can be effectively separated, the cell yield is high after gill digestion treatment, the cultured cells grow well, the physiological state is stable, the experimental research standard is achieved, the research approach of various disease problems possibly occurring in takifugu obscurus culture production can be fundamentally provided, and the bottleneck of research of the takifugu obscurus is broken through.

The invention relates to a cell sorting system based on endonuclease specific recognition. The cell sorting system based on the endonuclease specific recognition comprises the following components: a first single-stranded nucleotide, a second single-stranded nucleotide, an antibody, a cell sorting medium and a restriction endonuclease, wherein the first single-stranded nucleotide and the second single-stranded nucleotide are complementary, and a complementary region contains a recognition site of the restriction endonuclease; the first single-stranded nucleotide, the second single-stranded nucleotide, the antibody and the cell sorting medium are all modified, the modified first single-stranded nucleotide can be connected with the modified antibody, and the modified second single-stranded nucleotide can be connected with the modified cell sorting medium. The invention also provides a cell sorting method applying the cell sorting system based on the endonuclease specific recognition, and a kit and method for sorting DC (dendritic) cells, NK (natural killer) cells and CIK (cytokine-induced killer) cells from cell samples. By adopting the cell sorting technology provided by the invention, multiple cells can be sorted at the same time, sorting of a large number of delicate cells can be carried out, and the cell yield and survival rate are high.

The invention discloses a method for extracting and collecting peripheral blood mononuclear cells. The method comprises the following steps: 1) collecting peripheral blood; 2) pretreating peripheral blood; 3) performing centrifuging; 4) extracting plasma; 5) performing plasma inactivation; 6) collecting mononuclear cell layers; 7) centrifuging the mononuclear cell layers; 8) performing cleaning; and 9) extracting mononuclear cells. The invention belongs to the technical field of cell extraction, and particularly provides a method for separating peripheral blood mononuclear cells, which is usedfor obtaining the peripheral blood mononuclear cells by centrifugal extraction of Ficoll lymphocyte separation liquid and is high in cell yield and high in activity.

The invention belongs to the technical field of biology, and particularly relates to a method for separating single cells in animals and plants. The method comprises the steps of adding an active solution and a separating enzyme into a biological sample, and soaking to obtain a soaked mixture, wherein the active solution comprises sodium chloride, potassium chloride, a lanthanum compound and 4-hydroxyethylpiperazine ethanesulfonic acid; filtering the soaked mixture with a filter membrane; soaking and stirring the filter membrane and the precipitate on the filter membrane in the active solutionto enable single cells and microorganisms to be resuspended in the active solution, and collecting a composite cell suspension; and putting glass beads into the composite cell suspension, oscillating, and collecting the single-cell suspension. The cell activity is maintained by adopting the active solution, and the gelatinous substance in the sample is separated by adopting the separating enzyme,so that the cell yield and the cell purity are improved.

The invention discloses a method for isolating peripheral blood mononuclear cells, which comprises the following steps: (1) mixing the peripheral blood with Duchenne's phosphate buffer to obtain diluted peripheral blood; placing the lymphocyte separation solution in a centrifuge tube and Store in shading; (2) Tilt the centrifuge tube containing the lymphocyte separation solution so that the liquid level of the lymphocyte separation solution is at an angle of 50°-70° to the horizontal plane; add the diluted peripheral blood to the lymphocyte along the side wall of the centrifuge tube. The mixed solution is obtained from the cell separation solution; the volume ratio of the diluted peripheral blood to the lymphocyte separation solution is 1-2:1; (3) centrifuge the centrifuge tube containing the mixed solution and take the buffy coat to obtain the peripheral blood mononuclear cell layer. The separation method is easy to operate and is suitable for the separation of a large number of samples; it can realize aseptic operation, and the obtained peripheral blood mononuclear cells are suitable for clinical application; the damage to the isolated peripheral blood mononuclear cells is small, and the cell yield is high. will not be disturbed.

The invention discloses a mouse pancreas infiltrated immune cell separation method which comprises the following steps: digesting pancreas tissue by using mixed digestive juice of collagenase IV, DNAenzyme I and RPMI1640, filtering by using a nylon filtering net of 100 [mu]m, performing density gradient centrifugation, collecting middle-layer cells after centrifugation, and analyzing cell yield,a survival rate and cell types. Results show that (3.0+ / -1.2)*10<5> cells can be collected from each mouse, the survival rate of separated cells is (96.7+ / -2.2)%, flow cytometry shows that the contentof CD45 positive T cells is (97.1+ / -3.5)%, the content of CD3 positive T cells is (61.2+ / -5.7)%, the contents of CD4 positive T cells is (49.1+ / -5.1)%, the content of CD8 positive T cells is (47.2+ / -5.3)%, and the content of CD19 positive B cells is (32.8+ / -4.3)%. The conclusion is that the method is simply, efficient and stable, a great number of cells of a high survival rate and high purity arecollected, and a stable and reliable method is provided for related research on separation of pancreas infiltrated immune cells.

The present invention relates to the field of micro fiuidics. Specifically, the present invention relates to a novel flow cell for the selective enrichment of target particles or cells from a fluid. The flow cell exhibits a novel design which greatly improves the target particle or cell yield. The invention also provides a micro fiuidic device, comprising the flow cell according to the invention.In another aspect, the invention relates to the use of a flow cell or a micro fiuidic device of the invention for the isolation of target particles or cells from a fluid sample. Finally, the inventionrelates to a method for the selective enrichment of target particles or cells from a fluid using the flow cell of the invention.

The invention discloses a method for systematically freezing and storing human umbilical cord tissues according to structure levels. The method comprises the following steps of (1) separation and freeze storage of umbilical cord amniotic membrane tissues; (2) separation and freeze storage of umbilical cord blood vessel tissues; (3) separation and freeze storage of umbilical cord Wharton's jelly tissues. The invention also provides an umbilical cord restoration method. The freeze storage method provided by the invention has the advantages that the umbilical cord amniotic membrane, blood vesselsand Wharton's jelly tissues are sequentially peeled according to the structure levels; then through treatment, the materials are put into liquid nitrogen for freeze storage; three tissues of the umbilical cord are respectively stored; different freeze storage and restoration methods are used according to the specificity of each tissue; the freeze storage restoration activity and cell yield are favorably improved. The invention provides a set of methods for systematically storing complete umbilical cord tissues of various types; important biological resources are provided for the study of thestem cells from the umbilical cord and can be used for building tissue or cell resource sample databases derived from the umbilical cord. The method of the invention provides a solid foundation and basis for completing the human genetic resource databases from the umbilical cord.