40 results about "Donor cell" patented technology

The present disclosure describes systems, devices, and methods for high-throughput electroporation of a donor cell. The systems can include a perforated membrane having one or more layers, each comprising a plurality of channels disposed through each layer. The systems can also include spacer arrays in contact with a perforated membrane to create a plurality of cell electroporation chambers for high-throughput cell electroporation.

Provided are methods for rapid multiplex HLA genotyping and transcript quantitation. In some embodiments, the presently disclosed methods include providing a. sample from the cell, tissue, or organ that has mRNA derived from an HLA gene product; reverse transcribing the mRNA to produce a. pool of cDNAs; amplifying the cDNAs that result from reverse transcription of HLA-specific mRNAs to produce a pool of HLA-specific cDNAs; and determining the sequence of each HLA-specific cDNAs, whereby the HLA status of the cell, tissue, organ, or subject is determined. Also provided are methods for genotyping donor cells, tissues, and / or organs meant for transplantation into recipients. In some embodiments, the methods can be used to identify the presence of absence of each of HLA-A, -B, -C, -DRB 1 / 3 / 4 / 5, -DQA1, and -DQB1 in a biological sample.

The present disclosure provides bovine animals and methods to create recipient bovine animals for spermatogenic stem cell transplantation / complementation through modulation of the NANOS2 gene. In one embodiment genome editing was used to create bovine animals with insertions or deletions (indels) that inactivate or otherwise modulate NANOS2 gene activity so that resulting male bovines lack functional germ cells yet retain functional testicular somatic cells, and bovine females are fertile. These bovine males can then be transplanted or complemented with donor cells capable of giving rise to cells of the spermatogenic lineage, or spermatogenic stem cells and used for breeding.

Disclosed are compositions and methods for preventing graft versus host disease (GVHD) or allograft rejection in subjects receiving donor cells. Also disclosed are methods enhancing regulatory T (Treg) cells for use in preventing GVHD. Also disclosed are methods of suppressing alloreactive donor cells in a subject receiving transplant donor cells that involves adoptive transfer of the treated Treg cells. Also disclosed are enhanced Treg cells produced by the disclosed methods that have been engineered to express chimeric antigen receptor (CAR) polypeptide cells.

The invention belongs to the technical field of biological medicines, and particularly relates to a membrane-coated oncolytic virus as well as a preparation method and application thereof. The oncolytic virus contains a GSDMD N-terminal expression cassette regulated and controlled by a heated shock promoter, and pyroptosis can be induced under exogenous mild thermal stimulation, so that the virus is promoted to be quickly released, and reinfection and amplification of adjacent tumor cells are realized. A bionic nano-vesicle (iNV) formed by a donor cell membrane modified by genetic engineering is used as an outer layer to coat a genetically engineered oncolytic virus (GOV) to obtain a membrane-coated oncolytic virus iNV-GOV, so that the membrane-coated oncolytic virus iNV-GOV simultaneously has immunocompatibility and active tumor recognition capability, is suitable for solid tumors such as pancreatic ductal adenocarcinoma (PDAC), hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma (IHCC) and the like, and can be used for preparing an anti-tumor drug. And the tumor volume can be obviously reduced in mouse in-vivo model experiments.

This invention discloses a gene editing system for constructing a porcine nuclear transplantation donor cell model of atherosclerosis with double gene mutations in AF and its applications. The invention provides a kit comprising plasmid pKG-U6gRNA (APOE-E2-gRNA2), a target sequence binding region as shown in nucleotides 3-22 of SEQ ID NO: 25 (FBN1-gRNA4), a target sequence binding region as shown in nucleotides 3-22 of SEQ ID NO: 26 (FBN1-gRNA6), and FBN1-mutant-ss163 as shown in SEQ ID NO: 27. The kit is used for: preparing recombinant cells; preparing porcine atherosclerosis models; preparing atherosclerosis cell models, atherosclerosis tissue models, or atherosclerosis organ models. The plasmid pKG-U6gRNA (APOE-E2-gRNA2) is transcribed to obtain sgRNA with a target sequence binding region as shown in nucleotides 1-20 of SEQ ID NO: 11. APOE‑E2‑gRNA2 This invention lays the foundation for developing pig models of atherosclerosis through somatic cell nuclear transfer animal cloning technology.

Immunotherapeutic Monoclonal Antibody Complexes or Conjugates (IMAC) comprising readily accessible antibodies designed and approved for clinical use are provided using a one-step method that combines killing of existing cancer cells in parallel with induction of long-lasting anti-cancer vaccination. Methods for their use, alone or in combination with cancer killer cells including intentionally mismatched donor T cells, NK cells concomitantly with additional anti-cancer or immune activating agents, or activation of patient's own immune system for personalized treatment of cancer and elimination of undesirable non-malignant cells are also provided. In addition, treatment method based on IMAC can be applied for in vivo vaccination against cancer using an existing malignant lesion as internal anti-cancer vaccine by engagement of patients antigen presenting cells for induction of long-lasting anti-cancer vaccination in situ against residual or recurrent disease.

The application discloses a kit for constructing an ATM gene mutation ataxia-telangiectasia model pig nuclear transfer donor cell. The application provides a kit comprising ATM-gRNA1 shown in SEQ ID NO: 16, ATM-gRNA4 shown in SEQ ID NO: 17 and NCN protein. The application also provides a method for preparing a recombinant cell: co-transfecting a pig cell with ATM-gRNA1, ATM-gRNA4 and NCN protein to obtain a recombinant cell. The recombinant cell is a recombinant cell with ATM gene mutation. The kit is used for: preparing a recombinant cell; preparing an ataxia-telangiectasia model pig; preparing an ataxia-telangiectasia cell model or an ataxia-telangiectasia tissue model or an ataxia-telangiectasia organ model. The application has great application value for research and development of ataxia-telangiectasia drugs and revealing the pathogenesis of the disease.

This invention discloses a gene editing system for constructing porcine nuclear transplantation donor cells for ALS models with SOD1 gene mutations and its applications. This invention also provides a method for preparing recombinant cells: SOD1- g RNA1 (target sequence binding region as shown in nucleotides 3-22 of SEQ ID NO: 18), SOD1- g RNA6 (target sequence binding region as shown in nucleotides 3-22 of SEQ ID NO: 19), SOD1-mutant-ss160 (shown in SEQ ID NO: 20), and NCN protein (Cas9 protein or a fusion protein containing Cas9 protein) were co-transfected into porcine cells to obtain recombinant cells. This invention utilizes CRISPR / Cas9 technology combined with ssODN homologous recombination technology to perform point mutation gene editing of the SOD1 gene, mimicking the natural pathogenesis and genetic characteristics of ALS, and obtaining single-cell clones with precise point mutations in the SOD1 gene. This lays the foundation for later breeding of ALS disease model pigs using somatic cell nuclear transfer animal cloning technology.

The invention discloses a rapid breeding method for generating larimichthys crocea sperms based on spermatogonial stem cell transplantation. The rapid breeding method comprises the following steps: step S1, performing whole cryopreservation on larimichthys crocea testis; s2, separating the spermatogonial stem cells of the pseudosciaena crocea; s3, dyeing the spermatogonial stem cells of the pseudosciaena crocea; step S4, treatment of nibea albiflora receptors; and S5, transplanting the dyed donor larimichthys crocea spermatogonial stem cells into the nibea albiflora receptor testis. According to the method, when the nibea albiflora acceptor is treated, the traditional busulfan concentration of 40 mg / kg, twice injection and 35 DEG C high-temperature assistance is optimized into 60 mg / kg, single injection and 22 DEG C normal-temperature culture, optimal parameters induced by busulfan are screened out through experiments, the operation process is simplified under the action of ensuring that endogenous germ cells of the acceptor are reduced and immunological rejection is reduced, and the method is suitable for large-scale popularization and application. The harsh requirement of a high-temperature environment is avoided, and meanwhile the field planting efficiency of donor cells in receptors and the survival rate of the receptors are improved.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating said genetic modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near a gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or a component or a transcriptional regulator of a MHC-I or MHC-II complex, wherein genetic modification comprises an insertion of a polynucleotide encoding a tolerogenic factor and / or survival factor. The universal donor cells may further comprise at least one genetic modification within or near a gene that encodes a survival factor, wherein said genetic modification comprises an insertion of a polynucleotide encoding a second tolerogenic factor and / or a different survival factor.

Immunotherapeutic Monoclonal Antibody Complexes or Conjugates (IMAC) comprising readily accessible antibodies designed and approved for clinical use are provided using a one-step method that combines killing of existing cancer cells in parallel with induction of long-lasting anti-cancer vaccination. Methods for their use, alone or in combination with cancer killer cells including intentionally mismatched donor T cells, NK cells concomitantly with additional anti-cancer or immune activating agents, or activation of patient's own immune system for personalized treatment of cancer and elimination of undesirable non-malignant cells are also provided. In addition, treatment method based on IMAC can be applied for in vivo vaccination against cancer using an existing malignant lesion as internal anti-cancer vaccine by engagement of patient's antigen presenting cells for induction of long-lasting anti-cancer vaccination in situ against residual or recurrent disease.

This invention relates to the application of mitochondrial formulations in the preparation of drugs for treating chronic obstructive pulmonary disease (COPD). The mitochondrial formulations contain active mitochondria isolated from healthy donor cells, with a particle size of 400–1000 nm, possessing normal mitochondrial membrane potential and ATP generation function, and capable of being internalized by mammalian COPD lung epithelial cells. In vitro cell experiments have confirmed that this mitochondrial formulation can repair CSE-induced mitochondrial dysfunction in lung epithelial cells, inhibit mPTP opening, reduce mtDNA leakage into the cytoplasm, thereby blocking the activation of the cGAS-STING signaling pathway and reducing the release of inflammatory factors such as IL-1β and TNF-α. In vivo experiments in COPD mouse models show that tail vein injection of the mitochondrial formulation can effectively accumulate in lung tissue, significantly improving lung pathological changes such as emphysema and collagen fiber deposition in model mice. This invention provides a novel strategy for COPD treatment based on repairing mitochondrial function and intervening in inflammatory signaling pathways.

A method of producing cells into which exogenous mitochondria have been introduced, the production method including culturing recipient cells using cell culture equipment in which mitochondria isolated from donor cells are coated on the culture surface, and incorporating the mitochondria into the recipient cells. Cells produced by the method, the cells having exogenous mitochondria introduced thereinto; a cell culture substrate usable in the aforesaid production method and a method for manufacturing the same; and a cell culture kit.

This document provides genetically modified cells compatible with multiple subjects, such as universal donor cells; and methods for generating said genetically modified cells. These universal donor cells contain at least one genetic modification within or near at least one gene encoding a survival factor, wherein the genetic modification includes the insertion of a polynucleotide encoding a tolerogenic factor. These universal donor cells may further contain at least one genetic modification within or near a gene encoding one or more MHC-I or MHC-II human leukocyte antigens or components or transcriptional regulatory factors of the MHC-I or MHC-II complex, wherein said genetic modification includes the insertion of a polynucleotide encoding a second tolerogenic factor.

A method of modifying copy (1) and copy (2) of the β2-microglobulin (β2M) gene of a porcine donor animal, genetically modified porcine donor animal wherein copy (1) and copy (2) of the β2M gene of the porcine donor animal genome are modified, and a method of producing a donor porcine donor animal tissue or organ for xenotransplantation, wherein copy (1) and copy (2) of the β2M gene of the cells of said donor porcine donor animal are modified. A method of humanizing a genetically engineered donor animal by modification of the donor animal's expressed native β2-microglobulin protein through genetic alterations of the endogenous β2M genes, and as a result, the newly expressed β2M protein of the donor animal would contain some or all of the amino acids of the orthologous human β2-microglobulin (hβ2M) protein, in identity and orientation. This results in immune-compatible xenotransplants between donor animals and human recipients.

The invention discloses an application of a mitochondria-rich cell disintegration body in treatment of myocardial ischemia reperfusion injury. The invention provides an application of a mitochondrial-rich cell disintegration body in preparation of a product for preventing and / or treating myocardial ischemia-reperfusion injury. The cell disintegration body is extracted from mitochondrial donor cells treated by cytochalasin D. The invention further provides a preparation method of the mitochondrial-rich cell disintegration body. The mitochondrial-rich cell disintegration body has ultrahigh mitochondrial loading capacity and functional activity and excellent in-vivo and in-vitro stability; the functional mitochondria can be efficiently delivered to a heart injury area, the myocardial protection effect is achieved by activating an endothelial cell autophagy pathway, the heart function is remarkably improved, the infarct area is reduced, and the good clinical application prospect is achieved.

The present application relates to the application of mitochondrial transplantation preparation in the preparation of a drug for treating non-alcoholic fatty liver disease, wherein the mitochondrial transplantation preparation comprises active mitochondria isolated from healthy donor cells, and has normal membrane potential and mitochondrial respiratory function. In vivo experiments on a non-alcoholic fatty liver mouse model prove that tail vein injection of the mitochondrial transplantation preparation of the present application can effectively reduce the weight of the mouse, improve the fatty liver condition of the mouse, reduce the accumulation of fat in the liver tissue, and at the same time restore the abnormal liver function of the non-alcoholic fatty liver mouse.

The invention discloses a method for efficiently enriching anti-blue ear and porcine reproductive capacity edited pig donor cells. The method comprises the following steps: introducing CRISPR-Cas9 plasmids targeting a BMP15 gene, CRISPR-Cas9 plasmids targeting a CD163 gene and alternative reporter plasmids edited by double genes into the pig donor cells; culturing the porcine donor cells to enable gene editing and reporter gene expression to occur; detecting and sorting cells with positive report gene expression so as to obtain enriched double-gene editing donor cells; the alternative reporter plasmid for double-gene editing comprises the following elements which are connected in sequence: a recognition sequence of a BMP15 gene editing target, a transcription terminator sequence, a recognition sequence of a CD163 gene editing target and a fluorescent reporter gene sequence. According to the invention, the alternative reporter plasmid for BMP15 and CD163 double-gene editing is constructed, and the editing condition of target sites of the BMP15 and CD163 genes of a cell genome can be converted into a detectable red fluorescence phenotype, so that the enrichment efficiency of anti-blue ear high-fecundity edited pig donor cells is improved, and the double-gene editing efficiency of the cells is effectively improved.

The application discloses application of a gene editing system in preparation of a spinal muscular atrophy model pig nuclear transfer donor cell with SMN1 gene mutation. The application provides application of SMN1-gRNA3 shown in SEQ ID NO: 16, SMN1-gRNA4 shown in SEQ ID NO: 17 and NCN protein in preparation of a kit. The application also provides a method for preparing a recombinant pig cell: SMN1-gRNA3, SMN1-gRNA4 and NCN protein are co-transfected into a pig cell to obtain a recombinant pig cell. The recombinant pig cell is a recombinant pig cell with SMN1 gene mutation. The kit is used for: preparing a recombinant pig cell; preparing a spinal muscular atrophy model pig; preparing a spinal muscular atrophy cell model or a spinal muscular atrophy tissue model or a spinal muscular atrophy organ model. The application has great application value for research and development of spinal muscular atrophy drugs and revealing the pathogenesis of the disease.

A method for producing, from a donor cell, a low-antigenic cell in which a rejection reaction is reduced in a case where the cell is allogeneically transplanted into a recipient, the method including: determining human leukocyte antigen (HLA) alleles for the donor cell and the recipient, respectively; specifying an HLA allele that is present in the donor cell but is not present in the recipient; and disrupting or modifying the specified HLA allele to obtain a cell population including a cell not expressing an HLA protein specific to the donor cell, in which the cell not expressing the HLA protein specific to the donor cell is the low-antigenic cell.

Compositions and methods of transplanting cells by grafting strategies into solid organs (especially internal organs) are provided. These methods and compositions can be used to repair diseased organs or to establish models of disease states in experimental hosts. The method involves attachment onto the surface of a tissue or organ, a patch graft, a “bandaid-like” covering, containing epithelial cells with supporting early lineage stage mesenchymal cells. The cells are incorporated into soft gel-forming biomaterials prepared under serum-free, defined conditions comprised of nutrients, lipids, vitamins, and regulatory signals that collectively support stemness of the donor cells. The graft is covered with a biodegradable, biocompatible, bioresorbable backing used to affix the graft to the target site. The cells in the graft migrate into and throughout the tissue such that within a couple of weeks they are uniformly dispersed within the recipient (host) tissue. The mechanisms by which engraftment and integration of donor cells into the organ or tissue involve multiple membrane-associated and secreted forms of MMPs.

A system and method for collecting plasma comprises a control circuit configured to control operation of the system, the control circuit configured to receive one or more donor parameters. The control circuit estimates a physiological fluid amount of the donor based at least in part on the one or more donor parameters and calculates a target amount of plasma product comprising raw plasma and anticoagulant by multiplying a prestored constant by the estimated physiological fluid amount. The control circuit controls the system to operate draw and return phases to process whole blood into plasma product until a measured amount of plasma product in the collection container meets the target amount of plasma product.

This invention discloses a gene editing system for constructing porcine nuclear transplantation donor cells for a metoprolol reductase deficiency model with SPR gene mutations and its applications. The invention provides a kit comprising SPR-gRNA1 (SEQ ID NO: 16), SPR-gRNA4 (SEQ ID NO: 17), and NCN protein. The invention also provides a method for preparing recombinant cells: co-transfecting porcine cells with SPR-gRNA1, SPR-gRNA4, and NCN protein to obtain recombinant cells. The recombinant cells are recombinant cells with a mutated SPR gene. The kit is used for: preparing recombinant cells; preparing porcine metoprolol reductase deficiency models; and preparing metoprolol reductase deficiency cell models, tissue models, or organ models. This invention has significant application value for the development of drugs for metoprolol reductase deficiency and for elucidating the pathogenesis of this disease.

The present invention relates to donor cells engineered for use in general donor cell therapy, for allogeneic, ready-to-use administration, and with greater persistence in the immune system of a host. The invention also relates to the use of miRNA expression constructs in such engineered donor cells.

The application discloses an exosome drug loading method, which has two methods: (1) endogenous loading method: first, let the donor cells load drugs, and then secrete drug-containing exosomes; (2) exogenous loading method: after extracting the exosomes, the drugs are introduced by physical or chemical methods. The exosome drug loading method provided by the application can efficiently load the drugs into the exosomes and keep the activity of the drugs. The method is suitable for large-scale production.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating said genetic modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or component or transcriptional regulator of the MHC-I or MHC-II complex, at least one genetic modification that increases the expression of at least one polynucleotide that encodes a tolerogenic factor, and optionally at least one genetic modification that increases or decreases the expression of at least one gene that encodes a survival factor.

The application discloses a gene editing system for constructing a progeria model pig nuclear transfer donor cell with a ZMPSTE24 gene mutation and application thereof. The application provides a kit comprising ZMPSTE24-gRNA2 shown in SEQ ID NO: 16, ZMPSTE24-gRNA4 shown in SEQ ID NO: 17 and NCN protein. The application also provides a method for preparing a recombinant pig cell: co-transfecting ZMPSTE24-gRNA2, ZMPSTE24-gRNA4 and NCN protein into a pig cell to obtain a recombinant pig cell. The recombinant pig cell is a recombinant cell with a ZMPSTE24 gene mutation. The kit is used for: preparing a recombinant pig cell; preparing a progeria model pig; preparing a progeria cell model or a progeria tissue model or a progeria organ model. The application has great application value for the research and development of progeria drugs and the revelation of the pathogenesis of the disease.

The application discloses a method for preparing a breast cancer model pig nuclear transfer donor cell with a BRCA2 gene mutation and a special gene editing system thereof. The application provides a method for preparing a recombinant pig cell: BRCA2-gRNA1, BRCA2-gRNA2 and NCN protein are co-transfected into a pig cell to obtain a recombinant pig cell. The recombinant pig cell is a recombinant cell with a BRCA2 gene mutation. The application provides a kit comprising BRCA2-gRNA1 shown in SEQ ID NO: 16, BRCA2-gRNA2 shown in SEQ ID NO: 17 and NCN protein. The kit is used for: preparing a recombinant pig cell; preparing a breast cancer model pig; preparing a breast cancer cell model or a breast cancer tissue model or a breast cancer organ model. The application has great application value for research and development of a breast cancer drug and revealing the pathogenesis of the disease.

A method of use of a synthetic SCGB, and compositions thereof, to prevent the development or reduce the development of antibodies against a foreign antigen or set of antigens in the subject that receives the antigen(s) is provided. A synthetic SCGB being recombinant human CC10 protein is provided. The foreign antigen(s) is / are a cell, tissue or organ from a donor subject is administered, introduced, or transplanted into a recipient subject, a lung transplant, transplanted kidney, heart, liver, hematopoietic cell, or any other tissue or organ, antigen(s) is / are from human or non-human origin, purified therapeutic proteins or other drugs, food or drink, self-antigen or an allergen.