49 results about "Biological immortality" patented technology

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 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.

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 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.

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 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 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.

The invention relates to the field of cell engineering, and particularly discloses a preparation method and application of a conditionally immortalized human myocardial fibroblast model. According to the method, a Tet-on system driven by a cardiac fibroblast specific promoter is constructed to regulate and control a lentiviral vector of an SV40LT-tsA58 temperature-sensitive mutant gene, and after primary human cardiac fibroblasts are transduced, conditional immortalization of the cells is realized in the presence of an inducer doxycycline. The model can be stably amplified under an induction condition, the immortalized gene expression is closed after an inducer is removed, the primary physiological state of cells is recovered, and the high-fidelity response capability to fibrosis promoting and anti-fibrosis drugs is reserved. According to the method, the in-vitro proliferation limit of primary cells is broken through, the problems of species difference and ethics are avoided, and a humanized, high-fidelity and high-controllability standardized tool is provided for myocardial fibrosis pathological mechanism research and drug screening.

This invention discloses an engineered mitochondria, its preparation method, and its application. The preparation method of the engineered mitochondria includes: constructing immortalized mesenchymal stem cells; isolating and extracting mitochondria from the immortalized mesenchymal stem cells; pretreating the immortalized mesenchymal stem cells with IFN-γ and TNF-α to induce apoptosis, and extracting engineered apoptotic extracellular vesicle membranes; mixing the mitochondria with the engineered apoptotic extracellular vesicle membranes, and obtaining engineered mitochondria with their surface coated with the engineered apoptotic extracellular vesicle membranes after sonication and centrifugation. This effectively improves the stability and targeting of the engineered mitochondria, not only solving the problem of low cell uptake efficiency during direct mitochondrial transplantation, but also enhancing the functional recovery effect of mitochondria in damaged cells, providing a more efficient and stable source of mitochondria for engineered mitochondrial transplantation therapy.

The invention provides a Col4a5 + / -mouse immortalized podocyte line, and the preservation number of the Col4a5 + / -mouse immortalized podocyte line is CGMCC (China General Microbiological Culture Collection Center) NO.46563. The cell line has relatively strong stability guarantee and can be subjected to continuous passage for 30 generations. The Col4a5 < + / -> mouse immortalized podocyte line corresponds to the phenotype of a female XLAS carrier with a light disease degree, can be used for precise pathological modeling, realizes COL4A5 gene dose effect simulation, and can be used as a cell model for screening a treatment drug or a diagnosis drug for hereditary nephropathy. The cell line is subjected to immortalization treatment, can be massively cultured and subcultured, has relatively simple culture requirements, can obtain a large number of experimental cells in a short time, has a short total experimental period, is suitable for popularization, and overcomes the technical problems of difficulty in constructing a single allele mutation (Col4a5 + / -) cell line, low transfection efficiency, allele loss and the like in the prior art.

The invention relates to a method for evaluating ferroptosis sensitivity of NF2-related vestibular nerve sheath tumor Schwann cells. The method comprises the following steps: constructing NF2-mutated immortalized Schwann cells; carrying out ferroptosis induction and intervention experiments, and establishing a plurality of experiment groups; detecting ROS level, Fe < 2 + > concentration, GSH content, MDA level and protein expression of GPX4 and the like of each group of cells; sample preparation is carried out through a TEM method, and mitochondrial volume and morphological characteristics of each group of cells in a ferroptosis state are observed; detecting the cell viability, cell membrane damage and apoptosis proportion of each group of cells; and evaluating the influence of NF2 deletion on ferroptosis sensitivity. According to the method, a ferroptosis cell modeling scheme in which Erastin (an inducer) and Fer-1 (an inhibitor) are jointly intervened is designed, and a multi-dimensional ferroptosis evaluation system is combined, so that the stability and the result reproducibility of NF2 related vestibular nerve sheath tumor Schwann cell ferroptosis state evaluation are remarkably improved.

This invention provides a highly active recombinant human type III collagen, the amino acid sequence of which is shown in SEQ ID NO.2. The invention also provides a gene encoding the above-mentioned recombinant human type III collagen, a recombinant expression vector containing this gene, and an engineered Pichia pastoris strain containing the recombinant expression vector. Furthermore, this invention provides a method for preparing the above-mentioned recombinant human type III collagen and its applications. The advantages of this recombinant human type III collagen are: small molecular weight, high expression level, and excellent activity. It not only exhibits excellent proliferation, adhesion, and migration-promoting effects on human fibroblasts and immortalized human epidermal cells, but also possesses good hemostatic activity, making it widely applicable in hemostasis, wound repair, and medical aesthetics.

The invention discloses an immortalized goat bone marrow-derived macrophage for expressing an SV40LT gene and application of the immortalized goat bone marrow-derived macrophage, and belongs to the technical field of bioengineering. The immortalized goat bone marrow-derived macrophage provided by the invention is obtained by knocking an immortalized gene SV40LT into a primary goat bone marrow-derived macrophage ROSA26 site through a CRISPR-Cas 9 system, and the immortalized goat macrophage provided by the invention is an immortalized goat macrophage cell strain which is suitable for being infected with salmonella and brucella and can be continuously and stably passaged; the method can be effectively applied to immunological research on goat brucella infection resistance and production of brucella vaccines.

The invention discloses an immortalized egg shell gland epithelial cell line and application thereof. The invention belongs to the technical field of biology, and particularly relates to an immortalized egg shell gland epithelial cell line and application thereof. The construction method of the immortalized egg shell gland epithelial cell line comprises the following steps: 1) obtaining SV40 overexpression lentivirus; 2) transfecting the lentivirus obtained in the step 1) to chicken primary eggshell gland epithelial cells; amplifying the transfected cells for 3-4 generations, and screening puromycin to obtain successfully transfected eggshell gland epithelial cells; and (3) carrying out subculture on the successfully transfected eggshell gland epithelial cells obtained in the step (2) to obtain the stably subcultured immortalized eggshell gland epithelial cells. The immortalized egg shell gland epithelial cell provided by the invention can provide a reliable cell model for researching related functions of poultry egg shell glands, and has an important value for developing a novel feed additive capable of improving chicken genital tract health and egg shell quality.

The invention relates to the technical field of bioengineering, and provides a recombinant I-type collagen alpha2 chain protein and a preparation method thereof, and the amino acid sequence of the recombinant I-type collagen alpha2 chain protein is as shown in SEQ ID NO.3 or SEQ ID NO.4. The invention also provides a coding gene of the recombinant I-type collagen alpha2 chain protein, an expression vector, a recombinant pichia pastoris engineering bacterium and an application of the recombinant I-type collagen alpha2 chain protein. The recombinant I-type collagen alpha2 chain protein has the advantages that the activity is excellent, the proliferation and migration effects are achieved in human immortalized epidermal cells, and the recombinant I-type collagen alpha2 chain protein can be widely applied to the fields of medical cosmetology, cosmetics and the like.

The application provides a healthy human umbilical cord blood-derived immortalized NK cell line SZ093 and a preparation method and application thereof. The healthy human umbilical cord blood-derived immortalized NK cell line SZ093 is preserved in the Guangdong Provincial Microbial Culture Collection Center on September 26, 2025, and the preservation number is GDMCC No: 67036. The healthy human umbilical cord blood-derived immortalized NK cell line SZ093 provided by the application can still maintain good proliferation ability and high survival rate in a long-time (more than 1 year) culture process, and has high killing ability to tumor cells.

The invention relates to an immortalized cell line of human dermal papilla cells as well as a construction method and application of the immortalized cell line. The human dermal papilla cells are named as DYZ-20, the immortalized cell line is preserved in the China Center for Type Culture Collection on July 2, 2025, survival is detected, and the preservation number is CCTCC NO: C2025204. The immortalized cell line of the human dermal papilla cells can normally express alkaline phosphatase and a dermal papilla characteristic gene, and hair can grow after the immortalized cell line is transplanted into mouse skin.

The invention discloses an in-vitro stable subculture construction method of a yak ovarian granular cell line. Relates to the technical field of cell culture. Comprising the following steps: carrying out subculture and amplification on primary yak ovarian granular cells to obtain yGCs; and carrying out pCI-neo-hTERT plasmid transfection and establishment of a monoclonal cell. The immortalized yGCs constructed by using an hTERT plasmid method can provide infinitely proliferated homogenized cells, reduce damage to yaks and avoid individual differences, can retain the core function of regulating follicular development of yGCs, provides a high-quality model for analyzing molecular mechanisms such as seasonal oestrus and low reproduction efficiency of yaks, and has theoretical and application values.

The invention discloses a construction method of an immortalized chicken follicle granule cell line. The method specifically comprises the following steps: constructing an SV40LT gene overexpression lentiviral vector system, packaging and preparing a high-titer lentivirus; after chicken follicular granular cells are separated, lentivirus is used for transduction and puromycin resistance screening, and finally, an immortalized cell line is obtained through subculture. The constructed immortalized cell line has the following characteristics: continuous and stable passage can be realized, the biological functions of normal granular cells are reserved, and marker protein FSHR is specifically expressed; compared with primary cells, the proliferation capacity is remarkably enhanced, and the cell aging level is remarkably reduced. The method disclosed by the invention is simple and convenient to operate and high in efficiency, provides a reliable in-vitro model for research on follicular development, ovulation mechanism, reproductive endocrine regulation and the like of chicken, has important theoretical research and application values, and meanwhile, has a wide application prospect in reproductive development research related to reproduction in the poultry industry.

The invention discloses a multi-effect cell repair energy ointment and a preparation method thereof. The multi-effect cell repair energy ointment is prepared from 10-30 parts of a plant repair extract compound, 5-15 parts of coenzyme Q10, 3-10 parts of lecithin, 40-70 parts of vaseline, 5-12 parts of a natural moisturizing factor and 1-5 parts of a stabilizer. Wherein the plant repairing extract is used for removing active oxygen inside and outside cells, remarkably activating endogenous antioxidant pathways of the cells, inhibiting proinflammatory signals and creating a low-stress repairing microenvironment for skin cells; the coenzyme Q10 maintains energy metabolism of mitochondria; lecithin stabilizes cell membrane and immune cell structures, so that the immune cells keep higher activity under stress conditions. An in-vitro cell experiment verifies that the product can remarkably improve the survival rate of human immortalized keratinocytes and effectively promote the migration ability and the anti-oxidation defense function of human skin fibroblasts, and the multiple effects of promoting healing, relieving inflammation and enhancing the self-repairing ability of the skin are proved on the cellular level.