33 results about "Cell immortalization" patented technology

The present invention provides methods for immortalizing precursor cells that are non-terminally differentiated cells such as stem cells, said methods comprising culturing the precursor cells in the presence of a Notch agonist and one or more growth factors that support the proliferation but not differentiation of the non-terminally differentiated cells. The present invention further provides methods to induce the differentiation of immortalized cells, comprising growing the cells in the presence of a Notch agonist and at least one growth factor which supports the differentiation of the cell into a more specialized cell type. The immortalized and / or differentiated cells of the invention can be used to repopulate cell populations that have been diminished, for example as a result of infection or exposure to certain drugs. The invention further provides a cell culture comprising a population of non-terminally differentiated cells immortalized by the methods of the present invention and kits comprising reagents that promote the immortalization of precursor cells. The invention further provides methods for screening for Notch modulators and for identifying genes involved in processes of cellular differentiation.

The present invention provides methods for immortalizing precursor cells that are non-terminally differentiated cells such as stem cells, said methods comprising culturing the precursor cells in the presence of a Notch agonist and one or more growth factors that support the proliferation but not differentiation of the non-terminally differentiated cells. The present invention further provides methods to induce the differentiation of immortalized cells, comprising growing the cells in the presence of a Notch agonist and at least one growth factor which supports the differentiation of the cell into a more specialized cell type. The immortalized and / or differentiated cells of the invention can be used to repopulate cell populations that have been diminished, for example as a result of infection or exposure to certain drugs. The invention further provides a cell culture comprising a population of non-terminaly differentiated cells immortalized by the methods of the present invention and kits comprising reagents that promote the immortalization of precursor cells. The invention further provides methods for screening for Notch modulators and for identifying genes involved in processes of cellular differentiation.

An amino acid sequence Arg-Lys-Met-Leu-Lys-Ser-Thr-Arg-Arg-Gln-Arg-Arg (SEQ ID NO:1) functions as a protein transduction domain (PTD) and is capable of delivering small molecules, proteins, and nucleic acids to an intracellular compartment of a cell. An amino terminal lysine linker improves the efficiency of the PTD. A nuclear localization signal can be used to target the PTD to a cell's nucleus. The PTD can be used in PTD-cargo moiety complexes that can reversibly immortalize cells and increase cell viability in culture.

The instant invention provides methods for immortalizing cells. The invention further provides immortalized cell lines, e.g., neuronal cell lines, and methods of using these cell lines in screening assays.

The present invention relates to a method and to vectors for the immortalisation of cells independent of their type. It further relates to a cell or a cell line produced with the method or the vectors of the invention. The invention also relates to the use of this cell or cell line in in vitro applications and in the treatment of disease.

The invention discloses a construction method of an immortalized feeder layer cell strain, the immortalized feeder layer cell strain and application of the immortalized feeder layer cell strain. At least two immortalized genes are selected to be infected into feeder layer cells, and the two immortalized genes achieve a synergistic promotion effect on feeder layer cell immortalization through different cell immortalization promotion ways, and the immortalized feeder layer cell strain is obtained, so that the blank of a feeder layer cell immortalization method is filled, and wide cell resources are provided for evaluating the effect of treating tumors by drugs or controlling the proliferation of tumor cells and determining the proliferation or aging mechanism of the cells.

Reference controls for use with pharmacogenomic testing, and methods for their identification, preparation, and use, are disclosed. The reference controls can confirm that pharmacogenomic testing correctly identifies individuals that do or do not have the mutation of interest, in both clinical trial and patient treatment settings. The reference controls can be selected to include one or more mutations to be identified, and prescreened to confirm that they bind to one or more of the primers used in the pharmacogenomic testing. The reference controls are human genomic DNA that includes certain identified polymorphisms (mutations) of interest, ideally derived from individuals, pre-selected and optionally properly consented, which have one or more of the polymorphism(s) of interest. The reference controls can be prepared by targeted pre-screening of human patients, by examining the genotype or genetic profile of the patients, isolating cells with the desired mutation, optionally immortalizing the cells, and obtaining DNA from the cells. The prescreening of prospective donors can be targeted based on any of a number of factors, such as genes of interest, mutations within the genes of interest, and membership in a specific ethnic or disease state population. The genomic DNA can be pre-screened for its ability to be detected, using a standard pharmacogenomic test, as including a specific mutation. Examples of mutations of interest include those present in a Phase I or Phase II metabolic enzyme such as CYP2D6, CYP2C19, CYP2C9, CYP2C8, and CYP3A5, CYP3A4, CYP2A6, CYP2B6, UGT1A1, DPD, ERCC1, MDR1, ADH2, NAT1 and NAT2 or any other metabolic or disease gene.

Provided are a conditional suicide anti-tumour-targeting immune cell and a preparation method therefor. The preparation method comprises the steps of: A. using a recombinant DNA technique to insert a cell-immortalized gene into a cloning site of a first cloning vector and preparing same into a virion; B. selecting specifically an immune cell in human peripheral blood by using magnetic beads, and then infecting same with the virion prepared in step A to obtain an immortalized immune cell; C. using the recombinant DNA technique to insert a suicide gene into a second cloning vector, and then transferring same into the obtained immortalized immune cell to obtain a conditional suicide immune cell; D. using the recombinant DNA technique to insert a single-chain antibody gene into a cloning site of a third cloning vector and preparing same into a virion, and then transfecting the conditional suicide immune cell to obtain a conditional suicide anti-tumour-targeting immune cell.

The invention discloses an immortalized cell line of lamb testis Sertoli cells and its establishment method and application. The lamb testis Sertoli cell immortalized cell line is named as lamb testis Sertoli cell immortalized cell line hTERT‑LSC, and its preservation number is CCTCC NO: C2018202. The experiment found that the hTERT-LSC cell line can be continuously passaged for more than 60 generations, and its growth and proliferation characteristics remain unchanged. The titer of sheeppox virus propagated by this cell line is equivalent to that of lamb testicular Sertoli cell LSC, and the cell division rate is higher than that of LSC, and the division rate can reach 1:3. At the same time, the immortalized cell line is used to prepare the vaccine, which simplifies the production process, shortens the production cycle, reduces the production cost, and ensures the stable quality of the prepared sheep pox virus vaccine. Therefore, the proposal of the present invention provides a new technical means for the large-scale production of the sheep pox virus vaccine.

The invention discloses an immortalized cell line of neonatal bovine testis Sertoli cells, its establishment method and application. The immortalized Sertoli cell line of newborn bovine testis is named as the immortalized Sertoli cell line of newborn bovine testis hTERT‑NBSC, and its preservation number is CCTCC NO: C2018201. Experiments have found that hTERT-NBSC can be continuously passaged for more than 65 passages, and the growth and proliferation characteristics remain unchanged. The cell line is more sensitive to ORFV inoculation, can multiply ORFV virus efficiently, and can ensure the uniformity and stability of the virus. The virus titer of ORFV propagated by this cell line is equivalent to that of Sertoli cells of newborn bovine testis, and the cell division rate is higher than that of NBSC, and the division rate can reach 1:3. At the same time, the passage cell line is used to prepare the vaccine, which simplifies the production process, shortens the production cycle, reduces the production cost, and ensures the stable quality of the prepared sheep contagious impetigo vaccine. Therefore, the proposal of the present invention provides a new technical means for the large-scale production of the sheep contagious impetigo vaccine.

The invention discloses a fully humanized anti-hepatitis B virus monoclonal antibody as well as a preparation method and an application thereof, and belongs to the technical field of biomedicine and genetic engineering antibody. The monoclonal antibody comprises a heavy chain variable region and a light chain variable region; the amino acid sequence of the heavy chain variable region is shown as SEQ ID NO. 2; the amino acid sequence of the light chain variable region is shown as SEQ ID NO: 4; the coding DNA of the antibody comprises a heavy chain variable region and a light chain variable region, wherein the coding DNA sequence of the heavy chain variable region is shown as SEQ ID NO. 1; and the coding DNA sequence of the light chain variable region is shown as SEQ ID NO: 3. According to the method, B cells are immortalized through EBV virus, the natural fully-humanized hepatitis B virus monoclonal antibody is screened by combining a limited dilution method, and the screening method does not need a large expensive instrument, is simple and easy to implement and is suitable for being carried out in a conventional biology laboratory.

The invention discloses a method for lengthening, proliferating and culturing in-vitro telomeres of cartilage cells and human tissue engineering regenerated cartilage. The method and the human tissue engineering regenerated cartilage have the advantages that telomerase mRNA [(messenger RNA (ribonucleic acid)) transfection is carried out on the cartilage cells in an in-vitro manner, accordingly, the telomeres in cartilage cells obtained by means of culturing in follow-up procedures can be lengthened, proliferation and the viability of the cartilage cells and cartilage cells of elderly patients in particular can be obviously improved by the aid of synergistic effects of the telomerase mRNA transfection and small molecule compounds such as vitamin C, B18R or p65i, rapamycin and resveratrol in complete media, and the method and the human tissue engineering regenerated cartilage have important significance in improving or treating cell function decline due to senescence when the actually applied to regeneration medical treatment; the telomeres of the cartilage cells are not continuously or even frequently lengthened by the aid of the method for lengthening, proliferating and culturing the in-vitro telomeres of the cartilage cells, accordingly, genome insertion mutation risks or cell immortalization tumor formation risks can be prevented, the safety and the applicability can be greatly improved, and the method and the human tissue engineering regenerated cartilage can be effectively applied to the regeneration medical treatment.

The invention discloses a method for lengthening, proliferating and culturing in-vitro telomeres of cartilage cells and human tissue engineering regenerated cartilage. The method and the human tissue engineering regenerated cartilage have the advantages that telomerase mRNA [(messenger RNA (ribonucleic acid)) transfection is carried out on the cartilage cells in an in-vitro manner, accordingly, the telomeres in cartilage cells obtained by means of culturing in follow-up procedures can be lengthened, proliferation and the viability of the cartilage cells and cartilage cells of elderly patients in particular can be obviously improved by the aid of synergistic effects of the telomerase mRNA transfection and small molecule compounds such as vitamin C, B18R or p65i, rapamycin and resveratrol in complete media, and the method and the human tissue engineering regenerated cartilage have important significance in improving or treating cell function decline due to senescence when the actually applied to regeneration medical treatment; the telomeres of the cartilage cells are not continuously or even frequently lengthened by the aid of the method for lengthening, proliferating and culturing the in-vitro telomeres of the cartilage cells, accordingly, genome insertion mutation risks or cell immortalization tumor formation risks can be prevented, the safety and the applicability can be greatly improved, and the method and the human tissue engineering regenerated cartilage can be effectively applied to the regeneration medical treatment.

The invention provides methods and compositions for expanding cells that are not abundant or are difficult to obtain in pure form in culture, are in short supply (e.g., human cells), or have brief lifetimes in culture, using fusion polypeptide. The fusion polypeptide has a first region having the transport function of herpesviral VP22 protein or human immunodeficiency virus (HIV) TAT protein, and a second region with a polypeptide having cell immortalization activity, a polypeptide having telomerase-specific activity, or a polypeptide having telomerase gene activation activity. The resulting cells of the invention are suitable for use in cell therapy.