73 results about "Biological immortality" patented technology

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 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 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 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 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.

A DNA immortalization construct includes a first nucleic acid sequence derived from the 5′ end of human CDKN2A gene exon 2. An EF1α promoter sequence is connected to the first sequence. A first LoxP sequence is connected to the EF1α promoter sequence. A human TERT gene sequence is connected to the first LoxP sequence. A second LoxP sequence is connected to the human TERT gene sequence. An internal ribosomal entrance site (IRES) sequence is connected to the second LoxP sequence. An antibiotic selection gene sequence is connected to the IRES sequence. An SV40 poly-A signal sequence is connected to the antibiotic selection gene sequence. A second nucleic acid sequence is connected to the SV40 poly-A signal sequence and derived from 3′ end of human CDKN2A gene exon 2.

The invention relates to a CRISPR (clustered regularly interspaced short palindromic repeats)-mediated immortalized sheep fetal skin fibroblast line and application thereof in the field of variation or genetic engineering. The construction of the CRISPR-mediated immortalized sheep fetal skin fibroblast line comprises the following steps: step 1, constructing a Cas9 site-directed knockout plasmid containing an FGF5 gene target sequence; step 2, constructing a homologous repair plasmid Donor-2 kb HA-SV40LT containing a homologous arm of the FGF5 gene; step 3, constructing a donor DNA (Deoxyribose Nucleic Acid) containing an SV40LT protein expression cassette; and step 4, preparing the sheep fetal fibroblast line capable of stably expressing the SV40LT protein at a fixed point. The immortalized SFF-FGF5 / SV40LT monoclonal cell strain created by the invention can become an ideal tool cell line for genome editing research such as sheep CRISPR library screening, multiple gene editing, mutant gene function exploration and the like.

The application discloses a method for improving the passaging times of a mammary epithelial cell line and application thereof, and belongs to the technical field of mammary epithelial cell lines. The application first finds that the exogenous fragment HS4-SAR fragment in the attachment area of the insulator-scaffold is connected with a p-Ef1a-SV40LT-Puro vector or a pEF1a-eGFP vector, and the transfection of commercial immortalized cell lines and primary cell lines can significantly improve the passaging times of the mammary epithelial cell lines, the cell morphology is still normal after 120 generations, the malignant transformation does not occur, the gene expression level does not decrease, and the immortalization is realized; the established goat mammary epithelial cell line still has the ability of synthesizing and secreting proteins; the method for improving the passaging times of the mammary epithelial cell line is simple and easy to operate, has a significant effect on the immortalized cell lines and the primary cell lines, and is expected to improve the stable passaging times of various mammary epithelial cell lines, and reduce the cell cost of related research.

The invention discloses a human immortalized fallopian tube fibroblast strain as well as a construction method and application thereof, and belongs to the technical field of cell immortalization construction. The preparation method comprises the following steps: firstly, separating, culturing and purifying fibroblasts from human fallopian tube tissues by virtue of an enzyme digestion method, identifying markers of the fibroblasts, then introducing a simian virus 40 large T antigen gene into primary fibroblasts by virtue of a lentiviral vector, and screening by virtue of puromycin to obtain a cell strain capable of stably expressing SV40T. Finally, the human immortalized fallopian tube fibroblast which can be stably passaged is constructed. The cell strain has the characteristics of strong multiplication capacity and high hereditary stability, and the cells are not obviously aged along with the increase of the number of generations of the cells, which indicates that the human immortalized fallopian tube fibroblast line is successfully established. According to the invention, the problem that the in-vitro culture life of primary fibroblasts is limited is solved, and a reliable in-vitro cell model is provided for research in the fields of fallopian tube related diseases, ovarian cancer origin, tumor microenvironment and the like.

The application discloses a spinal muscular atrophy quality control product and a preparation method and application thereof. The application aims at the deficiency that the existing method of using Epstein-Barr virus to immortalize lymphocytes cannot efficiently transform spinal muscular atrophy lymphocytes into immortalized cells, and provides a transformation culture medium and a method for constructing immortalized spinal muscular atrophy lymphocytes by using the culture medium. By using the method, 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 immortalized spinal muscular atrophy lymphocytes are constructed by using the method, the required time is short, the success rate is high, the genetic background of the obtained immortalized cells is stable, the interoperability with clinical samples is high, the compatibility is good, the immortalized cells can be stably preserved, and the immortalized cells can be used as positive quality control products in gene detection of spinal muscular atrophy, so that the accuracy of detection can be ensured.

The application relates to the technical field of biology, and discloses a method for inducing a cell line of immortalized musk deer scent glands to synthesize dihydrotestosterone, wherein the cell line is obtained by transfecting a primary musk deer scent gland epithelial cell with a lentivirus carrying an SV40 Large T antigen and performing resistance screening; the cell line is in an epithelial cell form and stably expresses keratin 18, cholesterol side chain cleavage enzyme, 17 alpha-hydroxylase and 3 beta-hydroxysteroid dehydrogenase. Through lentivirus-mediated SV40 Large T antigen transfection, the primary musk deer scent gland epithelial cell breaks through the limit of replicative senescence and obtains unlimited proliferation capacity, effectively overcomes the defects that the primary cell is prone to fibrosis and rapidly loses metabolic function during in-vitro culture, provides a standardized cell model with a clear genetic background and persistent metabolic activity for biological manufacturing of dihydrotestosterone, and relieves the dependence on endangered animal live resources.

This invention discloses a method for constructing and applying a marine fish muscle stem cell line. The culture medium uses basal medium as the background medium and contains LIF growth factor, EGF growth factor, and FGF2 protein derived from the leopard-gill sea bass. The final concentration of FGF2 protein from the leopard-gill sea bass is 20-30 ng / mL, 30-45 ng / mL, 45-55 ng / mL, or 55-70 ng / mL. Passaging using this medium can construct a stable, rapidly dividing, and short-passaging immortalized marine fish muscle stem cell line, capable of stable passage for more than 50 generations. This provides raw materials for exploring the function and gene regulation mechanisms of myoblasts, and provides in vitro verification experimental conditions for further research on the effects of marine fish muscle stem cells on growth rate, laying the foundation for the breeding of superior marine fish species.

Provided herein is an immortalized MSC that heterologously expresses p53-upregulated-regulator-of-p53-levels (PURPL) lncRNA. Also provided is a method for making an immortalized MSC comprising heterologously expressing PURPL lncRNA in an MSC. Also provided are methods for treating a disease in a subject in need thereof comprising administering to the subject the immortalized MSC described herein or made by the methods described herein.

This invention relates to an immortalized chicken skeletal muscle satellite cell line, its construction method, and its applications, belonging to the field of biotechnology. The immortalized chicken skeletal muscle satellite cell line Im MSLC of this invention, with accession number CCTCC NO: C2023115, was deposited on July 5, 2023, at the China Center for Type Culture Collection, located at No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province, Wuhan University. The immortalized chicken skeletal muscle satellite cell line of this invention maintains a similar growth rate and consistent growth trend to primary chicken skeletal muscle cells after 10 passages, and retains the same cell morphology as primary cells after 22 passages; moreover, the immortalized chicken skeletal muscle satellite cell line still possesses differentiation ability after 10 passages. The immortalized chicken skeletal muscle satellite cell line provided by this invention reduces the use of chicken embryos and lowers the possibility of difficulty in obtaining stable and reliable results due to different batches of chicken embryos from which skeletal muscle cells are derived.