105 results about "Diploid cells" patented technology

A system and method for determining the genetic data for one or a small set of cells, or from fragmentary DNA, where a limited quantity of genetic data is available. Genetic data for the target individual is acquired and amplified using known methods, and poorly measured base pairs, missing alleles and missing regions are reconstructed using expected similarities between the target genome and the genome of genetically related subjects. In accordance with one embodiment of the invention, incomplete genetic data from an embryonic cell is reconstructed using the more complete genetic data from a larger sample of diploid cells from one or both parents, with or without genetic data from haploid cells from one or both parents, and/or genetic data taken from other related individuals. In accordance with another embodiment of the invention, incomplete genetic data from a fetus is acquired from fetal cells, or cell-free fetal DNA isolated from the mother's blood, and the incomplete genetic data is reconstructed using the more complete genetic data from a larger sample diploid cells from one or both parents, with or without genetic data from haploid cells from one or both parents, and/or genetic data taken from other related individuals. In one embodiment, the genetic data can be reconstructed for the purposes of making phenotypic predictions. In another embodiment, the genetic data can be used to detect for aneuploides and uniparental disomy.

Disclosed herein is a system and method for increasing the fidelity of measured genetic data, for making allele calls, and for determining the state of aneuploidy, in one or a small set of cells, or from fragmentary DNA, where a limited quantity of genetic data is available. Genetic material from the target individual is acquired, amplified and the genetic data is measured using known methods. Poorly or incorrectly measured base pairs, missing alleles and missing regions are reconstructed using expected similarities between the target genome and the genome of genetically related individuals. In accordance with one embodiment of the invention, incomplete genetic data from an embryonic cell are reconstructed at a plurality of loci using the more complete genetic data from a larger sample of diploid cells from one or both parents, with or without haploid genetic data from one or both parents. In another embodiment of the invention, the chromosome copy number can be determined from the measured genetic data of a single or small number of cells, with or without genetic information from one or both parents. In another embodiment of the invention, these determinations are made for the purpose of embryo selection in the context of in-vitro fertilization. In another embodiment of the invention, the genetic data can be reconstructed for the purposes of making phenotypic predictions.

Disclosed herein is a system and method for increasing the fidelity of measured genetic data, for making allele calls, and for determining the state of aneuploidy, in one or a small set of cells, or from fragmentary DNA, where a limited quantity of genetic data is available. Poorly or incorrectly measured base pairs, missing alleles and missing regions are reconstructed using expected similarities between the target genome and the genome of genetically related individuals. In accordance with one embodiment, incomplete genetic data from an embryonic cell are reconstructed at a plurality of loci using the more complete genetic data from a larger sample of diploid cells from one or both parents, with or without haploid genetic data from one or both parents. In another embodiment, the chromosome copy number can be determined from the measured genetic data, with or without genetic information from one or both parents.

A system and method for determining the genetic data for one or a small set of cells, or from fragmentary DNA, where a limited quantity of genetic data is available, are disclosed. Genetic data for the target individual is acquired and amplified using known methods, and poorly measured base pairs, missing alleles and missing regions are reconstructed using expected similarities between the target genome and the genome of genetically related subjects. In accordance with one embodiment of the invention, incomplete genetic data is acquired from embryonic cells, fetal cells, or cell-free fetal DNA isolated from the mother's blood, and the incomplete genetic data is reconstructed using the more complete genetic data from a larger sample diploid cells from one or both parents, with or without genetic data from haploid cells from one or both parents, and/or genetic data taken from other related individuals.

Disclosed herein is a system and method for increasing the fidelity of measured genetic data, for making allele calls, and for determining the state of aneuploidy, in one or a small set of cells, or from fragmentary DNA, where a limited quantity of genetic data is available. Poorly or incorrectly measured base pairs, missing alleles and missing regions are reconstructed using expected similarities between the target genome and the genome of genetically related individuals. In accordance with one embodiment, incomplete genetic data from an embryonic cell are reconstructed at a plurality of loci using the more complete genetic data from a larger sample of diploid cells from one or both parents, with or without haploid genetic data from one or both parents. In another embodiment, the chromosome copy number can be determined from the measured genetic data, with or without genetic information from one or both parents.

The invention relates to a culture medium for a normal epithelial cell of a human or mammal, primary separation culture, subculture, 3D gas-liquid culture and 3D matrigel culture methods, the normal epithelial cell generated by using the culture medium and the culture methods and application of the normal epithelial cell to a toxicological evaluation system. The culture medium is prepared by mixing DMEM and Ham's F-12NUTRIENT MIX according to the volume ratio of 3:1 and also adding 4-6% of fetal calf serum, 1-3nM triiodothyronine, 0.4-0.65% of insulin-transferrin-selenium reagent, 4-6mu g/ml transferrin, 9-11ng/mL epidermal growth factors, 0.3-0.5mu g/mL hydrocortisone, 0.5-1.5nM cholera toxin, 0.4-0.6mu g/mL amphoterrible B, 35-45mu g/mL gentamicin, 45-55nM calpeptin, 35-45ng/ml recombinant human IL-1RA and 3mu g/ml recombinant human R-Spondin-1. The culture medium disclosed by the invention can be used for carrying out separation culture or subculture on the normal epithelial cell of the human or the mammal and any other various tissue source, rapidly proliferating the normal epithelial cell in vitro and establishing a cell line; and the normal epithelial cell is a normal diploid cell and is applied to the toxicological evaluation system of the human or the mammal.

The present invention discloses preparation process of inactivated rotavirus vaccine. Rotavirus strains G1, G2, G3 and G4 are made to adapt human diploid cell line and propagate on human diploid cell line to obtain the producing strain of inactivated rotavirus vaccine, with the human diploid cell line being KMB17. The human diploid cell line used as the cell matrix for producing inactivated rotavirus vaccine is safer compared with available cell matrixes. Therefore, by means of the established technological platform, it is possible to develop inactivated rotavirus vaccine with other serotypes of rotavirus as the target.

The invention relates to the field of biotechnology, in particular to a virus seed for producing vaccines for preventing human rabies by utilizing human diploid cells (KMB17) and a preparation method thereof. In the invention, a rabies fixed virus CTN-1V5 strain is continuously subcultured in the human diploid cells (KMB17), and a terminal dilution method is used for screening viruses with highertiter, thereby obtaining a rabies virus strain which is suitable for the human diploid cells (KMB17) and has good immunogenicity and heredity stability, and culturing a rabies vaccine virus seed (CTN-DK strain) capable of efficiently reproducing in the human diploid cells (KMB17). By using the virus seed for producing a human diploid cell (KMB17) rabies vaccine, the risk caused by residual heterogonous DNA (deoxyribonucleic acid) in the vaccine which is currently used at home can be effectively avoided, and the safety and practicability of the rabies vaccine in China are further improved, thus the invention has great social and economic benefits.

The invention relates to the preparative way of a kind of freeze-drying varicella vaccine, which has something to do with the biotechnology. The main making procedures include the following steps: the amplification and passage of Varicellavirus strain Oka to the MRC-5 strain in human's diploid cells; the harvest of the vaccine liquids; freeze-drying. The main property of the vaccine is that it adopts 0.5-1.2%(g/ml) gelatine, 0.6-1.2%(g/ml) glutamic sodium, 0.3-0.9%(g/ml) carbamide and 0.05-0.2%(g/ml) arginine as the freeze-drying protective agents, the previous concentrations stand for the final concentration of the protective agents. The good quality of the vaccine lies in that: the freeze-drying protective agents are developed by the producers themselves to make sure the stability of the vaccine. The result of storage stability test at 2-8DEG C shows that: there's no evident difference between the samples that stored for 18 months and the new-produced samples.

The invention discloses a human diploid cell rabies vaccine and a preparation method thereof, and belongs to the field of vaccines. A CDKHBP-1 strain is inoculated into a human diploid cell WI-38, and the rabies vaccine is obtained through separation and purification. The human diploid cell does not contain any contaminants or oncogenicity, and is obvious superior to an animal cell serving as a medium for producing the vaccine; and the method is suitable for large-scale industrial production, a purification process is perfect, and the vaccine contains a few impurities and has high purity and immunogenicity.

A purified diplontic cell vaccine in the form of freeze-dried powder injection or liquid injection for rabies is prepared through inoculating rabies virus into human diploied cells.

The invention discloses a method for detecting the titre of live viruses, which comprises the following steps: firstly, infecting diploid cells or other adherent cells with dilute viruses to be detected, culturing and fixing the infected cells; secondly, adding virus antiserum (first antibody) into the fixed cells, culturing and washing, adding a second antibody marked by horseradish peroxidase (HRP), culturing and washing, adding coloring solution for color development and producing red or brown spots, or adding virus antiserum marked by the HRP into the fixed cells, culturing the virus antiserum and washing, adding the coloring solution for color development and producing red or brown spots; thirdly, calculating the virus titre according to the quantity of the red or brown spots and the dilution multiple, or calculating the 1gTCID50 value of the viruses according to the dilution degree and the hole numbers of the spots. The detection method can be applicable to the titre detection of various live viruses, can shorten the detection time of the titre of the viruses due to fast detection speed, has good specificity as the immune spots are caused by special immune reaction, low detection cost as well as simple and convenient operation and does not need special instruments of optical or fluorescent microscopes, and the like or other reagents.

The invention provides a method of preparing a hepatitis A inactivated vaccine, which comprises the following steps of inoculating mixed and absorbed hepatitis A virus strain SH and a human embryo diploid cell MRC-5 to hepatitis A virus propagated in a cell factory, digesting the cell in the virus propagation peak to obtain cell virus liquid, removing impurity proteins of the cell by ultrasonication, chloroform extraction and ultrafiltration through ultrafiltration membranes, degerming and filtering by gel filtration chromatography and purification, and absorbing by an aluminium hydroxide adjuvant so that the hepatitis A inactivated vaccine is prepared. The result of in vivo effectiveness experiments performed on a mouse shows that the hepatitis A inactivated vaccine prepared by the method of the invention has higher ED50 and better immunogenicity than the contract strain. The method of the invention can improve the safety of the vaccine, simplify the technique, shorten the productionperiod, reduce the production cost, and realize the scale production of the hepatitis A inactivated vaccine.

The invention relates to a preparation method for diploid cell Japanese encephalitis vaccine and purified encephalitis vaccine. The method uses a serum-free medium to culture human diploid cell Japanese encephalitis purified vaccine, thus greatly reducing anaphylactic reaction and being beneficial to purification.

The invention discloses a method for improving a survival rate of human diploid cells. The method comprises the following steps: digesting the cells by using a trypsin solution, centrifuging, and removing supernatant; suspending the cells by using a frozen stock solution, wherein a solvent of the frozen stock solution is normal saline or TC199 serum-free medium, and each milliliter of solvent contains 3 percent of human albumin, 5 percent of dimethyl sulfoxide, 0.5 percent of sodium carbonate, 2 percent of hydroxyethyl starch, 5 percent of serine and 6-10 percent of dimethylacetamide. According to the preservation method provided by the invention, the survival rate of the human diploid cells can be improved.

The invention provides a protein-free serum-free medium suitable for diploid cell culture and a preparing method thereof. The medium contains calcium chloride, magnesium sulfate, potassium chloride, sodium dihydrogen phosphate, L-arginine hydrochloride, L-asparaginate, L-cystine dihydrochloride, L-histidine hydrochloride, L-leucine, L-isoleucine and the like. The medium can meet the requirement of the growth or maintenance stage of diploid cells and prevents contact with exogenous viruses possibly existing in protein and serum to reduce potential safety hazards; as the components are simple and definite, standard products can be purchased in the market, the difference between different product batches is small, and the step of purifying down-stream products is simplified; a scientific basis is provided for large-scale standard production. In addition, the protein-free serum-free medium can be applied in the growth or maintenance stage of the diploid cells.

The invention discloses a method for producing a novel freeze-dried live attenuated varicella vaccine. The method is characterized in that a human diploid cell is used as a culture medium, a reactor culture process is applied to inoculate a varicella OKa strain, and an invented freeze-drying protective additive containing sorbitol and recombinant human serum albumin is used to produce the freeze-dried live attenuated varicella vaccine. By applying the reactor culture technology, the production cost is reduced, the defects of low yield, large production occupied area, high labor intensity and the like of a static culturing process can be overcome, the yield of vaccines can be increased, and the virus titer stability can be improved; furthermore, the protective additive containing sorbitol and recombinant human serum albumin is used and does not contain gelatin, so that animal derived ingredients can be removed, the content of toxin in the vaccine can be remarkably reduced, the stimulation and hazard of the vaccine on a human body can be greatly reduced, and the safety and stability of the vaccine can be improved.

The invention discloses an application of human embryonic lung fibroblast strains Walvax-2 in the preparation of a rabies inactivated vaccine. The human embryonic lung fibroblast strains Walvax-2 are susceptible to rabies viruses, and when the human embryonic lung fibroblast strains are used for producing the rabies virus vaccine, the diploid cell rabies virus inactivated purified vaccine for people can be obtained, wherein the vaccine is high in antigen purity, good in immune effect, high in safety and low in price.

The invention discloses a rapid adaptation method of rabies vaccine virus strains for the human body on diploid cells of the human body. Fresh rabies viruses are inoculated on the diploid cells of the human body, poisoned passage is carried out, only 4-5 cycles are required, passage is carried out for 12-15 passages, the virus titer rises above 7.0logLD50/ml of the P15 from 3.0logLD50/ml of the P1, the adaptation cycle of the viruses is shortened by at least 50%, on the premise that the virus immunogenicity is not affected, the virus yield is rapidly improved, and manpower and material resources are saved.

A vaccine of the encephalitis B virus for preventing viral encephalitis B is prepared through inoculating said virus into human diploid cells, culturing, naturalizing, adapting, amplifying, concentrating and purifying.

The invention discloses a method for cultivating triploid dendrobium huoshanense by a fusion breeding method of diploid cells and pollen cells. The method comprises the following steps: (1) selecting dendrobium huoshanense varieties from different producing areas, which are good in stress resistance, quick in growth, thick in stalks, long in stem height, various in tillering, low in fiber content and high in medicinal composition content as a triploid cultivating source plant parent; (2) performing aseptic seeding on source plant seeds in a solid culture medium to obtain protocorm; (3) performing sterile culture on the source plant pollen in the solid culture medium to obtain a pollen cell population; (4) preparing protoplasts of the pollen cells and the protocorm cells to perform cell fusion; (5) culturing the fused cells on the solid culture medium to differentiate the fused cells into young plants, and then detecting the triploid plants; and (6) performing tissue culture on the triploid plants to obtain a large number of triploid young plants.

The invention provides a method for preparing genotype F mumps attenuated live vaccine. The method comprises the following steps of inoculating mumps attenuated strain MHM-19 to human embryo lung diploid cell MHL-3 which is cultured to a monolayer; releasing virus through frozen-thawed cell in the peak of viral multiplication; centrifuging and removing cell debris; adding a protective agent to centrifuged virus suspension to obtain a semi-finished product of vaccine; and freezing and drying the semi-finished product to obtain the finished product of genotype F mumps attenuated live vaccine. After a rhesus monkey is injected with the prepared MHM-19 attenuated live vaccine for immunizing, good humoral immunity is induced to be produced, and the detection results obtained at different times after immunization show that the MHM-19 attenuated live vaccine has immunizing potency superior to that of reference mumps vaccine S79; in addition, the safety experiment shows that the rhesus monkey does not suffer from parotitis-related symptom after being injected with MHM-19 attenuated live vaccine, the pathological tissue nearly has no obvious difference with that of S79 strain by observing, so that the MHM-19 attenuated live vaccine has high safety.