125 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 discloses an RSV infection immortalized tree shrew aorta vascular endothelial cell model, and particularly relates to a method for separating and culturing primary aorta vascular endothelial cells from tree shrew aorta, further establishing an immortalized tree shrew aorta vascular endothelial cell line, taking the cell line as an RSV virus culture medium, and constructing an RSV infection immortalized tree shrew aorta vascular endothelial cell model. Establishing an RSV infected immortalized tree shrew aortic vascular endothelial cell model, and preparing a high-infection-titer RSV virus; the invention provides key support for preparation and detection of RSV virus and construction of a tree shrew in-vivo infection model, provides a stable virus source for development of comparative medical research or vaccine evaluation, and has far-reaching significance for deep exploration of RSV pathogenic mechanism, drug and vaccine evaluation and the like.

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 monoclonal goat intramuscular precursor fat cell immortalized cell line as well as a preparation method and application thereof, and relates to the field of immortalized cells. The method comprises the following steps: acquiring precursor fat cells in primary goat muscle; the method comprises the following steps: infecting precursor adipocytes in primary goat muscle with a lentivirus carrying SV40-T, and adding puromycin to screen positive cells infected with the lentivirus carrying SV40-T; a limited dilution method is used, the screened positive cells are laid on a plate according to one cell per hole, and the monoclonal goat intramuscular precursor fat cell strain is obtained after culture; and carrying out amplification culture on the monoclonal goat intramuscular precursor fat cell strain to obtain the immortalized cell line of the monoclonal goat intramuscular precursor fat cell. According to the immortalized cell line obtained by the method, while the morphology and the function of the precursor fat cells in the goat muscle are ensured, the passage generation number of the cells is remarkably increased, and the cells do not need to be repeatedly sampled and separated, so that the same genetic background of the cells is ensured, and the cost and the workload are greatly saved.

Provided are a liver precursor cell and a construction method therefor and the use thereof. The method for constructing liver precursor cells comprises the steps of: S0, providing liver precursor cells, a mixed solution and a reprogramming culture medium, wherein the mixed solution comprises a serum-free culture medium, polyethyleneimine and a CLP plasmid; S1, carrying out a transfection treatment on the liver precursor cells and the mixed solution to obtain transfected liver precursor cells; and S2, placing the transfected liver precursor cells into the reprogramming culture medium for subculture to obtain immortalized liver precursor cells. The method solves the problems that in the prior art, liver precursor cells cannot be transformed into immortalized cell strains by means of immortalizing genes and cannot be continuously passaged in vitro.

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 biology, and discloses a method for constructing immortalized thymoma primary epithelial cells. The method comprises the following steps: step 1, tissue dissociation and mixed cell population acquisition; step 2, performing differential liquid change to remove immune cells; step 3, immortalization treatment; step 4, carrying out monoclonal screening; step 5, amplification and verification; and step 6, passage and storage. The method is mainly used for establishing a stable immortalized cell line in a laboratory for subsequent scientific research. The specific application comprises tumor biology research; anti-tumor drug screening; cancer-related gene function research; and cell signal transduction mechanism research and the like. For tumors which do not have mature cell lines or are difficult to establish primary cell lines, such as thymoma, the invention is beneficial to improving the success rate of establishing immortalized tumor primary cell lines and promoting the development of subsequent basic scientific research.

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 provides a recombinant I-type collagen alpha1 chain polypeptide, the amino acid sequence of which is as shown in SEQ ID NO.2, and the recombinant I-type collagen alpha1 chain polypeptide contains two GER integrin recognition sites. The invention also provides a recombinant I-type collagen alpha1 chain polypeptide coding gene which is shown as SEQ ID NO.1 and is used for coding the recombinant I-type collagen alpha1 chain polypeptide. Meanwhile, the invention also provides a recombinant expression vector containing the recombinant I-type collagen alpha1 chain polypeptide coding gene, recombinant pichia pastoris engineering bacteria, and a preparation method and application of the recombinant I-type collagen alpha1 chain polypeptide. The recombinant I-type collagen alpha1 chain polypeptide designed by the invention is small in molecular weight, contains two GER integrin recognition sites, is excellent in activity, has a certain migration effect in human immortalized epidermal cells, and can be widely applied to the fields of medical cosmetology, cosmetics and the like.

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 purpose of the present invention is to provide a therapeutic agent for demyelination diseases, which comprises, as an active ingredient, a culture supernatant of immortalized mesenchymal stem cells into which a specific gene has been inserted. The present invention provides a therapeutic agent for demyelinating diseases, which contains, as an active ingredient, a culture supernatant of immortalized mesenchymal stem cells into which either an hTERT gene or a pTERT gene, and a bmi-1 gene, an HPV-E6 gene, or an HPV-E7 gene are inserted. The mesenchymal stem cells are preferably derived from a human or a pig, and the culture supernatant preferably contains a component that acts on the receptor c-MET of a hepatocyte growth factor (HGF), said component being obtained by culturing the mesenchymal stem cells for 24-96 hours.