94 results about "Biological immortality" patented technology

The invention belongs to the technical field of biology, and particularly relates to an immortalized alpaca kidney cell line and a construction method and application thereof. The immortalized alpaca kidney cell line disclosed by the invention is named as immortalized alpaca kidney cells (Alpaca kidney cells) BNalpaca 1.0, and the preservation number of the immortalized alpaca kidney cell line is CGMCC (China General Microbiological Culture Collection Center) No.46058. The construction method of the immortalized alpaca kidney cell line comprises the following steps: extracting and culturing alpaca primary kidney cells; infecting alpaca primary kidney cells by using SV40 large T antigen immortalized lentivirus; carrying out resistance screening on the immortalized alpaca kidney cells; carrying out alpaca kidney cell immortalized cell detection and identification; and obtaining the immortalized alpaca kidney cell line. The immortalized alpaca kidney cell constructed by the invention has the advantages of simple culture, repeated passage, good homogeneity, high expression quantity of constructed exogenous genes and the like, and provides a more powerful tool cell for research on alpaca immunity; the immortalized alpaca kidney cell line provides a new tool for virus infection mechanism, drug in-vitro function detection and antibody drug development and screening.

The invention belongs to the technical field of biomedicine, and discloses an immortalized cell line for human kidney suspected cell carcinoma as well as a culture method and application of the immortalized cell line. The immortalized cell line for the human kidney suspected cell carcinoma, disclosed by the invention, is named as a human kidney suspected cell carcinoma cell line Loya a-710 (Homo sapiens), the classification name of the Latin name of the immortalized cell line is Chromophore sensor cell carcino: Loya a-710, and the preservation number of the immortalized cell line is CCTCC (China Center for Type Culture Collection) NO: C2025123. The human kidney suspected cell carcinoma cell line Loya-710 disclosed by the invention is used as an in-vitro model to rapidly grow in tissue culture, and mitochondrial mutation characteristics, a vesicle structure and a classical immunohistochemical marker of ChRCC are reserved. The invention provides a precious tool for further research on ChRCC heredity, molecules and biological characteristics, and also provides a powerful new model for mitochondrial diseases.

The invention relates to the technical field of cell engineering, and particularly discloses a construction method and application of a sika deer immortalized renal epithelial cell line, and the construction method comprises the following steps: taking renal epithelial primary cells from healthy sika deer renal cortex tissues to obtain primary renal epithelial cells; treating the primary renal epithelial cells with trypsin to obtain a cell suspension; inoculating the cell suspension into a complete culture medium, and transfecting with lentivirus loaded with SV40 large T antigen genes to obtain transfected cells; 3 [mu] g / mL puromycin is applied to the transfected cells for selective culture, so that non-transfected cells are eliminated, and drug-resistant cells are obtained; carrying out continuous passage on the drug-resistant cells for at least 30 generations to obtain the immortalized renal epithelial cell line of the sika deer; according to the invention, through a specific action mechanism (inhibiting a p53 / pRb pathway and blocking cell cycle exit) of the SV40 large T antigen, a proliferation limit caused by inhibition of telomerase activity of primary cells of the cervidae animals is overcome, and a cell resource library capable of realizing continuous passage is established.

The invention discloses a preparation method and application of a conditioned culture solution for improving the development rate of bovine in-vitro embryos. The preparation method comprises the following steps: culturing immortalized bovine endometrial epithelial cells by using a DMEM / F12 cell culture solution containing 10% FBS, collecting the cell culture solution in the culture process, and filtering to obtain an immortalized bovine endometrial epithelial cell conditional culture solution, namely the conditional culture solution capable of improving the development rate of the bovine in-vitro embryos. The prepared conditioned culture solution can simulate a liquid environment of an in-vivo environment, is used for culturing in-vitro embryos such as parthenogenetic activated embryos and in-vitro fertilized embryos, improves the cleavage rate and blastocyst rate of the in-vitro embryos and improves the development rate of the in-vitro embryos of cattle.

The invention relates to the technical field of recombinant proteins, and relates to a recombinant XII type collagen as well as a preparation method and application thereof. The recombinant XII type collagen is formed by connecting 5-30 repetitive units in series, and the amino acid sequence of each repetitive unit is as shown in SEQ ID NO: 1. The nucleic acid molecule is used for coding the recombinant XII type collagen; a vector comprises the nucleic acid molecule. According to the invention, a nucleotide sequence for coding the amino acid sequence is inserted into a vector to obtain a recombinant plasmid vector, then the recombinant plasmid vector is introduced into a host cell for culture, and then separation and purification are carried out to obtain the recombinant XII type collagen. The recombinant XII type collagen provided by the invention is easy to obtain, can realize efficient secretory expression, and meets the requirements of industrial production; the biological activity is good, and the proliferation and migration of human immortalized cartilage cells can be promoted.

The invention discloses a combined evaluation method for in-vitro and in-vivo red fading and relieving effects of cosmetic raw materials, which comprises the following steps: in-vitro model evaluation: inducing a human immortalized umbilical vein endothelial cell model by adopting TNF-alpha, and detecting the secretion amount of a prostacyclin I2 factor to obtain a score of an in-vitro model; in-vivo model evaluation: adopting a chick embryo chorioallantoic membrane model, and obtaining a score of the in-vivo model according to the degree of vasoconstriction caused by the to-be-detected raw material sample acting on the chick embryo chorioallantoic membrane; and comprehensive evaluation: based on the score of the in-vitro model and the score of the in-vivo model, comprehensively evaluating the red fading and relieving efficacy of the raw material sample to be tested. According to the technical scheme, the defect that in the prior art, no cosmetic raw material soothing efficacy evaluation method for the vascular reactivity increasing mechanism exists is overcome, efficacy evaluation is more accurate, the method has the advantages of being comprehensive, short in test period, easy to operate, economical and convenient, and an effective reference basis is provided for screening and compounding schemes of red fading and soothing raw materials.

The invention discloses a human normal kidney immortalized cell line NRC-X1 and application thereof, and belongs to the field of microbial animal cell lines. The human normal kidney immortalized cell line is named as NRC-X1, and the preservation number is CCTCC (China Center For Type Culture Collection) NO: C2025168. The invention also discloses the application of the human normal kidney immortalized cell line NRC-X1 in a virus infection mechanism. The invention also discloses application of the human normal kidney immortalized cell line NRC-X1 in research of drug renal toxicity evaluation. The invention also discloses the application of the human normal kidney immortalized cell line NRC-X1 in bioartificial kidney and cell therapy. The invention also discloses application of the human normal kidney immortalized cell line NRC-X1 in research of kidney physiological and pathological mechanisms.

The invention discloses a panda immortalized kidney cell line GPK-1 as well as a preparation method and application thereof, and belongs to the technical field of immortalized cells. The preparation method comprises the following steps: (1) collecting kidney tissues of dead pandas, and soaking the kidney tissues in a protective solution; (2) carrying out two-step digestion on the kidney tissue treated in the step (1); and (3) carrying out culture passage on the digested cells, then infecting the passage cells with lentiviruses carrying neomycin resistance genes and SV40-LT genes, and then screening to establish the panda immortalized kidney cell line. The invention provides a technology for efficiently separating and culturing panda nephrogenic cells, and establishes an immortalized panda nephrogenic cell line GPK-1. Compared with primary kidney cells of pandas, the GPK-1 retains normal karyotype and related molecular characteristics and is remarkably improved in the aspects of cell passage times, proliferation efficiency and transfection efficiency.

The invention belongs to the field of biological medicine, and provides circular RNA for treating multiple endocrine tumor type I and application of the circular RNA. According to the invention, after different optimization and design are carried out on the circular RNA for coding Menin, the circular RNA with relatively good expression level and long half-life period is finally obtained, and the anti-tumor effect is mainly achieved by supplementing cell-deficient Menin. Compared with the prior art, the circular RNA for expressing the Menin protein provided by the invention is more stable in intracellular expression and longer in half-life period. In an immortalized parathyroid gland cell model, the Menin protein can be expressed by transfecting the circular RNA for 6 hours, and the circular RNA has good treatment potential for multiple endocrine tumor type I.

The invention discloses a yak intramuscular precursor fat immortalized cell line and a preparation method thereof. The method comprises the following steps: (1) obtaining yak primary intramuscular precursor fat cells; (2) performing SV40T lentivirus infection on the precursor fat cells in the yak muscle and screening cell strains: performing SV40T lentivirus infection by utilizing a 1 / 2 small-volume infection method to obtain immortalized cells after the precursor fat cells in the yak primary muscle are infected with SV40T, namely immortalized cells for short; puromycin is adopted to screen immortalized cells; and diluting the screened cells by using a limited dilution method, and carrying out enlarged culture to obtain the immortalized cell line. The yak intramuscular precursor fat immortalized cell line constructed by the invention has genetic homogeneity, the heterogeneity influence of cell populations is eliminated, and an in-vitro cell model is provided for subsequent research related to yak intramuscular fat cells.

The invention provides an immortalized NK cell line SZ093 derived from healthy human umbilical cord blood as well as a preparation method and application of the immortalized NK cell line SZ093. The immortalized NK cell line SZ093 derived from the healthy human umbilical cord blood is preserved in Guangdong Microbial Culture Collection Center on September 26, 2025, and the preservation number is GDMCC No: 67036. The immortalized NK cell line SZ093 derived from the healthy human umbilical cord blood, provided by the invention, can still keep good multiplication capacity and high survival rate in a long-time (more than one year) culture process, and has high killing capacity on tumor cells.

The invention discloses a method for increasing the passage frequency of a mammary epithelial cell line and application of the method, and belongs to the technical field of mammary epithelial cell lines. It is found for the first time that an insulator-stent attachment area exogenous fragment HS4-SAR fragment is connected with a p-Ef1a-SV40LT-Puro vector or a pEF1a-eGFP vector, commercial immortalized cell lines and primary cell lines are transfected, the passage number of mammary gland epithelial cell lines can be remarkably increased, and the mammary gland epithelial cell lines can be obtained. After 120 generations of passage, the cell morphology is still normal, malignant transformation is avoided, and the gene expression level is still not reduced, so that immortalization is realized; the established milk goat mammary gland epithelial cell line still has the capability of synthesizing and secreting protein; the provided method for increasing the passage frequency of the mammary epithelial cell line is simple and easy to operate, has a remarkable effect on an immortalized cell line and a primary cell line, and is expected to increase the passage frequency of stable passage of various mammary epithelial cell lines and reduce the cell cost of related research.

The invention belongs to the technical field of cell engineering, and particularly relates to a construction method and application of a mammal rapamycin target protein self-activated mouse model and a mouse embryo fibroblast line. According to the invention, serine (Serine, S) at site 2215 of mTOR protein is mutated into tyrosine (Tyrosine, Y), and a transgenic mouse model with activation mutation of mTORS2215Y, namely a mouse model with self-activation of mammal rapamycin target protein, is constructed. A mouse embryo fibroblast line which can be immortalized is constructed by utilizing the transgenic mouse model, so that the related mechanism research of mTOR self-activation mutation can be carried out at the cellular level, and the screening of related therapeutic drugs can be carried out.

The invention discloses a human immortalized pure renal cyst cell line SRC-X1 and application thereof, and belongs to the field of microbial animal cell lines. The human immortalized pure renal cyst cell line is named as SRC-X1, the cell line is preserved in the China Center for Type Culture Collection on September 19, 2025, and the preservation number is CCTCC NO: C2025303. The human immortalized pure renal cyst cell line SRC-X1 is applied to preparation, screening or evaluation of renal cyst treatment drugs. The invention also discloses the application of the human immortalized pure renal cyst cell line SRC-X1 as a renal cyst cell model. The human immortalized pure renal cyst cell line SRC-X1 is applied to renal cyst basic or clinical research as an experimental material.

Described herein is an immortalized mammalian cell configured for production of a bioproduct of interest, which cell has a safe harbor locus selected from an Adeno Associated Virus 1 (AAVS1) locus, a CCR5 locus, and a Rosa26 locus into which a Human Papillomavirus (HPV) E6 gene sequence and an HPV E7 gene sequence have been inserted. Also disclosed herein are methods for immortalizing a mammalian cell for production of a bioproduct of interest as well as methods of producing a bioproduct of interest.

The present invention belongs to the field of biomedical technology and discloses an immortalized cell line of human renal chromophobe carcinoma, a culture method and an application thereof. The immortalized cell line of human renal chromophobe carcinoma of the present invention is named human renal chromophobe carcinoma cell line Loya-710 (Homo sapiens), and its Latin name is Chromophobe renal cell carcinoma:Loya‑ 710 , deposited with CCTCC NO: C2025123. The human chromophobe renal cell carcinoma cell line Loya-710, as an in vitro model, grows rapidly in tissue culture and retains the mitochondrial mutations, vesicle structure, and classic immunohistochemical markers characteristic of ChRCC. This invention provides a valuable tool for further studying the genetic, molecular, and biological characteristics of ChRCC and offers a powerful new model for mitochondrial disease.

The invention discloses an immortalized rabbit liver cell line as well as a construction method and application thereof, and the construction method is characterized in that an exogenous oTERT gene is introduced into rabbit embryo liver cells, so that the rabbit embryo liver cell line is immortalized. The successfully immortalized rabbit embryo liver cell line can be stably subcultured for more than 10 generations, and does not have an aging phenomenon. Compared with a traditional method (introducing exogenous immortalized genes such as cancer suppressor genes p53 and SV40TAg into cells), the TERT-mediated cells belong to normal cells, the growth characteristics of the TERT-mediated cells are kept consistent with those of the normal cells, the TERT-mediated cells belong to immortalization in a real sense, and a scientific and stable cell material can be provided for subsequent RHDV research. The construction method is simple, easy to implement and easy to popularize.

The present disclosure relates to cultured tissue, methods for production of the cultured tissue, e.g., cultured meat, that is not genetically modified. Further, the disclosure provides method of temporary immortalization of cells for propagation in vitro.

The application discloses a kind of immortalization human umbilical cord mesenchymal stem cell NOTCH1-LNR overexpression stable cell line construction method and its application in liver differentiation, it is related to immortalized cell technical field.The application first connects target gene to vector, and the slow virus plasmid containing target gene is constructed, the slow virus plasmid containing target gene is introduced into 293T cell, and high titer slow virus containing target gene is generated;Immortalized human umbilical cord mesenchymal stem cell is infected with slow virus, and mixed clone cell strain is screened, and the immortalized human umbilical cord mesenchymal stem cell NOTCH1-LNR overexpression stable cell line is obtained.The application proves that the differentiation efficiency of immortalized human umbilical cord mesenchymal stem cell NOTCH1-LNR overexpression can effectively improve liver-like cell to glycogen staining and Western blotting experiment.This research result is expected to provide new technical thought for the next step development to improve the yield of MSCs liver differentiation.

The present application relates to a CRISPR-mediated immortalized sheep fetal skin fibroblast cell line in the field of variation or genetic engineering and application thereof.The construction of the CRISPR-mediated immortalized sheep fetal skin fibroblast cell line of the present application comprises the following steps: step one, constructing a Cas9 site-directed knockout plasmid containing an FGF5 gene target sequence; step two, constructing a homologous repair plasmid Donor-2 kb HA-SV40LT containing an FGF5 gene homologous arm; step three, constructing a donor DNA containing an SV40LT protein expression frame; and step four, preparing a sheep fetal fibroblast cell line stably expressing SV40LT protein.The immortalized SFF-FGF5 / SV40LT monoclonal cell strain created by the present application can become an ideal tool cell line for genome editing researches such as sheep CRISPR library screening, multiple gene editing and mutation gene function exploration.

The application discloses a kind of construction method and application of seawater fish muscle stem cell line, culture medium with basic medium as background medium;Culture medium includes LIF growth factor, EGF growth factor and leopard bristletooth-derived FGF2 protein;The final concentration of leopard bristletooth-derived FGF2 protein is 20~30 ng / mL, 30~45 ng / mL, 45~55 ng / mL or 55~70 ng / mL. Through the medium, immortalized seawater fish muscle stem cell line with good stability, cell division vigorous, short subculture time can be constructed, and can be stably subcultured for more than 50 generations. It provides raw materials for exploring the function of myoblasts, gene regulation mechanism, and provides the prerequisite for in vitro verification experiment for further exploring the influence of seawater fish muscle stem cells on growth rate, and lays a foundation for good strain breeding of seawater fish.

The application belongs to the field of microbial animal cell lines, and relates to a human immortalized simple renal cyst cell line SRC-X1 and an application thereof. The human immortalized simple renal cyst cell line is named SRC-X1, the cell line has been preserved in the China Center for Type Culture Collection on September 19, 2025, and the preservation number is CCTCC NO: C2025303. The application of the human immortalized simple renal cyst cell line SRC-X1 in preparation, screening or evaluation of renal cyst treatment drugs. The application of the human immortalized simple renal cyst cell line SRC-X1 as a renal cyst cell model. The application of the human immortalized simple renal cyst cell line SRC-X1 as experimental material in renal cyst basic or clinical research.

The present invention discloses a giant panda immortalized kidney cell line GPK‑1, a preparation method and a use thereof, and belongs to the field of immortalized cell technology. The preparation method is as follows: (1) collecting kidney tissue of a dead giant panda and immersing it in a protective solution; (2) performing two-step digestion on the kidney tissue treated in step (1); (3) culturing and propagating the digested cells, and then infecting the propagated cells with a lentivirus carrying a neomycin resistance gene and an SV40‑LT gene, and then screening and establishing a giant panda immortalized kidney cell line. The present invention provides a technology for efficiently isolating and culturing giant panda kidney-derived cells, and establishes an immortalized giant panda kidney-derived cell line GPK‑1. Compared with primary giant panda kidney cells, GPK‑1 retains normal karyotype and related molecular characteristics, and has been significantly improved in terms of cell passage times, proliferation efficiency and transfection efficiency.

The application provides a method for constructing an immortalized bovine nasopharyngeal tonsil epithelial cell line and application, and belongs to the technical field of cell engineering. The healthy yellow bovine nasopharyngeal tonsil is used as a material, and the primary epithelial cells are separated by enzyme digestion. The 3rd to 4th generation cells are taken, infected with SV40 T lentivirus (MOI=80), and screened by 1.5 μg / mL puromycin to obtain a stable immortalized cell line. The cell line is continuously passaged to the 50th generation and still stably expresses SV40LT protein and epithelial specific markers CD326, CD324 and KRT8, maintains a typical pavement stone-like morphology, normal proliferation and no malignant transformation. Infection experiments show that the cell is highly sensitive to FMDV, supports efficient replication, and the virus titer can reach 10 7.5 TCID 50 / mL. The application overcomes the defects of the existing immortalized bovine nasopharyngeal tonsil epithelial cell model, such as limited source, large batch difference, low sensitivity and poor persistence, and provides a stable and reliable in vitro model for the research of foot-and-mouth disease prevention and control related mechanisms, drug screening and vaccine development.

The application discloses immortalized chicken embryo epithelioid cells and a preparation method and application thereof, and the preparation method of the immortalized chicken embryo epithelioid cells comprises the following steps: extracting primary chicken embryo epithelial cells from a chicken embryo; and culturing the primary chicken embryo epithelioid cells in a conditional medium containing chicken Noggin protein for subculture, so as to obtain the immortalized chicken embryo epithelioid cells. The immortalized chicken embryo epithelioid cells prepared by the preparation method can be stably subcultured for a long time, can maintain stable proliferation ability after multiple subcultures, reduces individual differences, and improves the reliability and repeatability of experimental results. In addition, the preparation method does not introduce exogenous genes or change the genotype of the cells, ensures the stability at the gene level, and avoids the interference of the exogenous genes on the stability of the cell phenotype.

Described herein are systems, methods, and interconnecting porous hydrogel blocks for ex vivo kidney tissue generation. A block may comprise a 3D continuous polymeric matrix with a network of microporous cavities, and may be configured to interconnect with at least one other block. A system may comprise at least one block and be configured to cultivate one or more kidney-like organ components when seeded with at least one HEK293 cell and at least one kidney cell isolated from at least one specimen. A method may comprise seeding at least one block with at least one HEK293 cell and at least one kidney cell isolated from at least one specimen

The invention discloses a spinal muscular atrophy quality control product as well as a preparation method and application thereof. The invention provides a transformation culture medium and a method for constructing an immortalized spinal muscular atrophy lymphocyte by utilizing the transformation culture medium, aiming at the defect that the existing method for immortalizing the lymphocyte by utilizing EB (Epstein-Barr) virus cannot efficiently transform the spinal muscular atrophy lymphocyte into an immortalized cell. By utilizing the method disclosed by the invention, the immortalized spinal muscular atrophy lymphocyte strain can be constructed within 30 days, and the corresponding immortalized cell line can be obtained by subculturing the immortalized spinal muscular atrophy lymphocyte strain. When the method is used for constructing the immortalized spinal muscular atrophy lymphocyte, the required time is short, the success rate is high, and the obtained immortalized lymphocyte is stable in genetic background, high in intercommunity with a clinical sample, good in compatibility and capable of being stably stored, and can be used as a positive quality control material for gene detection of spinal muscular atrophy, so that the detection accuracy is ensured.

The present invention provides a low-molecular-weight human epidermal cell autophagy regulatory peptide DKX-9 and its applications. According to the general cell autophagy regulation mechanism and its theory, a small-molecule synthetic peptide is newly designed and synthesized, which can regulate the autophagy activity and effect improvement of human epidermal cells. The low-molecular-weight human epidermal cell autophagy regulatory peptide DKX-9 of the present invention adopts a peptide sequence composed of 9 amino acids of five types of amino acids (threonine, proline, tyrosine, asparagine, and arginine), and its amino acid composition and sequence are SEQ ID No.1: TPYNRRRRY-NH2. After applying it to human immortalized epidermal cells (HaCat), it is confirmed that it can up-regulate the expression levels of positive autophagy-related proteins Beclin-1, LC3-II, LAMP1, and TFEB, and can down-regulate the expression level of the negative autophagy-related protein p62. The autophagy regulatory peptide DKX-9 of the present invention has positive significance and medical value for future intervention in skin aging and prevention and treatment of certain skin diseases, and plays an important role in promoting the research and application of synthetic peptides in related disciplines such as cell autophagy in the future.

The application discloses a new use of androsta-4,6,8(9),13(14)-tetraen-3,11,16-trione, namely, application of the androsta-4,6,8(9),13(14)-tetraen-3,11,16-trione in preparation of a medicine for treating lymphoma. 50 The IC value of the androsta-4,6,8(9),13(14)-tetraen-3,11,16-trione in the SR cell line is detected by a CCK-8 method, and the cytotoxicity of the androsta-4,6,8(9),13(14)-tetraen-3,11,16-trione on human umbilical vein endothelial cells Huvce and human immortalized keratinocytes Hacat is detected, in the body, the androsta-4,6,8(9),13(14)-tetraen-3,11,16-trione significantly inhibits tumor growth and shows good biological safety and no systemic toxicity; TUNEL staining, gamma-H2AX staining and neutral comet experiment prove that the compound of the application inhibits the growth of SR by causing serious damage to DNA, the application not only adds the diversity of the biological activity of the androsta-4,6,8(9),13(14)-tetraen-3,11,16-trione, but also provides a new direction for developing a new medicine for treating lymphoma.

The invention belongs to the technical field of cell culture, and particularly relates to a construction method of an immortalized goat umbilical cord mesenchymal stem cell line. The method comprises the following steps: by taking newborn goat lamb umbilical cord tissues as a research object, cleaning goat lamb umbilical cords, stripping blood vessels, separating Wharton's jelly, cutting the Wharton's jelly into 1mm < 3 > tissues, culturing by using a DMEM / F12 complete culture medium containing fetal calf serum, carrying out passage, cryopreservation, repeated resuspension and centrifugation, and inoculating into a culture dish for culture; lentiviruses containing SV40 large T antigens are packaged in vitro and are transduced into goat umbilical cord mesenchymal stem cells, and immortalization is realized after long-term subculture. The result of the embodiment shows that the construction method of the immortalized goat umbilical cord mesenchymal stem cell line provided by the invention is strong in repeatability, simple and efficient in operation steps, good in stem cell growth state and stable in physiological state.