143 results about "Donor cell" patented technology

The invention belongs to the technical field of biology, and discloses a reagent combination or kit for constructing intestinal organs and application of the reagent combination or kit. According to the reagent combination or the kit, intestinal organs can be obtained from cell-derived epithelial cells obtained from a donor in a non-invasive manner, and the obtained intestinal organs can be cryopreserved and recovered and can be amplified in vitro for a long time; transcriptome characteristics are similar to those of real human small intestine tissues, and typical marker genes of various small intestine pedigree cell types are highly expressed; compared with intestinal organs obtained through induction of pluripotent stem cells, the intestinal organs have more intestinal pedigree characteristics, and the intestinal function development is more mature; after being promoted to be mature, the intestinal organ also highly expresses genes related to drug absorption and metabolism, has drug absorption capability similar to that of an immortalized intestinal cell line, but more prominently shows intestinal cell lineage characteristics, is closer to an intestinal environment in a real human body, and can be used for screening intestinal disease drugs; the intestinal barrier function is realized.

Disclosed are compositions and methods for preventing graft versus host disease (GVHD) in subjects receiving donor cells. In particular, chimeric antigen receptor (CAR) polypeptides are disclosed that can be used with adoptive cell transfer suppress alloreactive donor cells. Also disclosed are immune effector cells, such as T cells or Natural Killer (NK) cells, that are engineered to express these CARs. Therefore, also disclosed are methods of suppressing alloreactive donor cells in a subject receiving transplant donor cells that involves adoptive transfer of the disclosed immune effector cells engineered to express the disclosed CARs.

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 a survival factor, wherein the genetic modification comprises an insertion of a polynucleotide encoding a tolerogenic factor. The universal donor cells may further 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 said genetic modification comprises an insertion of a polynucleotide encoding a second tolerogenic factor.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating the 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.

Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating the genetically 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.

Several embodiments disclosed herein relate to methods and compositions for enhanced expansion of NK cells in culture, through the repeated co-culturing of NK cells with feeder cells and the selective use of stimulatory interleukins. In several embodiments, the methods utilize one or more soluble interleukins as culture media supplements at one or more time points during expansion of the NK cell, or other immune cell, which results in a highly expanded and highly cytotoxic population of cells, for use in, for example allogeneic cellular immunotherapy.

The invention discloses a rapid and efficient extraction method of extracellular vesicles rich in mitochondria. The extraction method comprises the following steps: (1) culturing mitochondrial donor cells in vitro; (2) treating the mitochondrial donor cells with cytochalasin D; (3) collecting the cells; (4) oscillating a cell suspension; according to the method disclosed by the invention, by combining cytochalasin D treatment with a subsequent gradient centrifugal separation step, a sample of extracellular vesicles rich in mitochondria can be efficiently extracted, meanwhile, the defects existing in a traditional mitochondrial transplantation technology are overcome, and a new thought and method are provided for clinical application of mitochondrial transplantation.

The invention discloses a conjugational transfer-assisted orthogonal targeted mutation evolution system, which comprises a donor strain and a receptor strain, a donor strain intracellular muter performs targeted mutation on a target sequence to obtain a mutation sequence, a non-self-transferable conjugational transfer functional plasmid is introduced into the donor strain, and a non-self-transferable conjugational transfer functional plasmid is introduced into the receptor strain. The mutant sequence is unidirectionally and horizontally transferred from a donor strain cell to a receptor strain cell, and the complete orthogonality of the mutants in the initial donor strain and the mutant sequence in the receptor strain is realized through the orthogonality between the donor strain and the receptor strain. The results of a sacB-sucrose reverse screening experiment show that the mutation frequency of an ortho-MutaT7 system reaches 2.96 * 10 <-2 > at 96 hours, which is 1840.9 times of the natural mutation frequency of the sacB in donor cells, and the ortho-MutaT7 system generates mutation of 12 basic group change types in a target sequence. The Ortho-MutaT7 system developed by the invention realizes mutation diversity and also has orthogonality, leakage mutation and off-target mutation of mutants in host cells are fully avoided, and a direct relationship between a mutation sequence and a phenotype is established.

Disclosed are compositions, in particular, organoid compositions, derived from more than one donor cell. Further disclosed are methods of making compositions, for example, organoid compositions, that comprise a differentiated cell population derived from more than one donor cell. Donor cells may include, for example, a precursor cell such as an embryonic stem cell or other precursor cell. The disclosed methods use synchronization conditions to produce a synchronized pooled-precursor cell population, which may then be differentiated into an organoid composition. Methods of using the compositions are also disclosed.

The invention discloses an InDel molecular marker combination and application of the InDel molecular marker combination in heterologous cell therapy pharmacokinetic detection. The InDel molecular marker combination is selected from one or more of 161 InDel loci on a human chromosome; relative to a reference genome hg19, position information and mutation types of the 161 InDel sites are shown in a table 1. The InDel molecular marker combination can be used for distinguishing donor cells and recipient cells for cell therapy, different donors and recipients are detected by adopting the InDel molecular marker combination, after a specific InDel site is selected, the mutation frequency of the InDel site of a sample (the recipients subjected to cell therapy) after medication is detected, and the mutation frequency of the InDel site is detected. According to the InDel site mutation frequency, the proportion of heterologous cells in the sample can be calculated, the problem that no mature analysis method exists in heterologous cell therapy pharmacokinetic research is effectively solved, and the InDel molecular marker combination can also distinguish donors and receptors with fewer sites.

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 a survival factor, wherein the genetic modification comprises an insertion of a polynucleotide encoding a tolerogenic factor. The universal donor cells may further 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 said genetic modification comprises an insertion of a polynucleotide encoding a second tolerogenic factor.

The application discloses a gene editing system for constructing a three-gene (GHR gene, IGF1 gene and IGF2 gene) combined mutation small pig nuclear transfer donor cell and application thereof. The application provides a kit comprising GHR-E5-gRNA2 shown in SEQ ID NO: 36, GHR-E5-gRNA3 shown in SEQ ID NO: 37, IGF1-E4-gRNA1 shown in SEQ ID NO: 38, IGF1-E4-gRNA2 shown in SEQ ID NO: 39, IGF2-E4-gRNA2 shown in SEQ ID NO: 40, IGF2-E4-gRNA7 shown in SEQ ID NO: 41 and NCN protein. The NCN protein is a Cas9 protein or a fusion protein with the Cas9 protein. The application adopts CRISPR / Cas9 technology combined with double gRNA editing to perform combined knockout of the GHR gene, the IGF1 gene and the IGF2 gene, and obtains a single cell clone with three-gene combined knockout, thereby laying a foundation for breeding small pigs and growth and development disorder or retardation model pigs through somatic cell nuclear transfer animal cloning technology in the later stage.

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.

To provide, generally, methods and compositions for transferring a genetic circuit from one prokaryotic cell ("donor cell") to another prokaryotic cell ("recipient cell" or "target cell" used interchangeably herein).SOLUTION: The present disclosure relates to: prokaryotic donor cells comprising (i) a genetic circuit of interest and (ii) one or more expressed transcriptional repressor proteins; and the use of the donor cells in efficient transfer of the genetic circuit into a prokaryotic recipient cell. The genetic circuit includes nucleic acid sequences encoding an RNA molecule or protein of interest.SELECTED DRAWING: None

The present disclosure relates to products and methods relating to transplant recipient antibodies bound to donor cell membrane fragments. Products and methods provided are used to separate and collect transplant recipient antibodies from donor cell membrane fragments, as well as to characterize the transplant recipient antibodies. The products and methods are useful for allotransplant or xenotransplant recipients.

The application discloses a gene editing system for constructing a muscle weakness disease model pig nuclear transfer donor cell with a DOK7 gene mutation and application thereof. The application provides application of DOK7-gRNA2 shown in SEQ ID NO: 16, DOK7-gRNA4 shown in SEQ ID NO: 17 and NCN protein in preparation of a kit. The application further provides a method for preparing a recombinant cell, comprising the following steps: co-transfecting a pig cell with DOK7-gRNA2, DOK7-gRNA4 and NCN protein to obtain a recombinant cell. The recombinant cell is a recombinant cell with a mutated DOK7 gene. The use of any of the above kits is to prepare a recombinant cell, to prepare a muscle weakness model pig, to prepare a muscle weakness cell model, a muscle weakness tissue model or a muscle weakness organ model. The application has great application value for research and development of muscle weakness disease drugs and revealing the pathogenesis of the disease.

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.

The present disclosure provides compositions and methods for cell transplantation therapy based on forced expression of an exogenous HLA-F protein in donor cells to be transplanted into a subject. In some embodiments, the donor cells express an exogenous chimeric HLA-F protein comprising an extracellular region comprising an HLA-F alpha 1 domain, an HLA alpha 2 domain, an HLA-F alpha 3 domain, a linker and a β2m protein.

To provide genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and to provide methods of generating the genetic modified cells.SOLUTION: The universal donor cells comprise at least one genetic modification within or near at least one gene encoding a survival factor, the genetic modification comprising the insertion of a polypeptide encoding a tolerogenic factor. The universal donor cell may further comprise 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 regulators of the MHC-I or MHC-II complexes, the genetic modification comprising the insertion of a polypeptide encoding a second tolerogenic factor.SELECTED DRAWING: Figure 1

The invention discloses a line establishment method and a culture system of bovine trophoblast stem cells. The stem cell line is obtained by separation and induction from bovine blastocyst. The embryonic stem cell line has stability. The method can be applied to various life science and medical fields such as bovine animal breeding and breeding, gene editing models, animal cloned donor cells, medical musculoskeletal injury experiments, drug development carriers, vaccine production cell banks, embryo quality analysis and the like, and can be applied to large-scale production and application.

A resource allocation method is applied to a communication system based on massive MIMO. The method includes: A donor eNodeB detects whether a target terminal exists in a donor cell, where a correlation between a wireless air interface channel of the target terminal and a wireless air interface channel of a relay node is greater than a target threshold; and if the target terminal does not exist, the donor eNodeB determines, for the relay node through resource spatial multiplexing, one or more beams used for data backhaul, where the one or more beams are beams formed by a massive antenna array of the donor eNodeB through air interface beamforming.

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.

Methods for improving engraftment of donor cells in a subject thereof are provided. Such methods can comprise providing cells that have been modified to express a first isoform of a target protein (e.g., C-X-C motif chemokine receptor 4 (CXCR4)), administering the donor cells to the subject, and then selectively inhibiting host cells in the subject based on their expression of the second isoform of the target protein, thereby improving engraftment of donor cells in the subject. Also provided are combinations for administration to a subject in need thereof, wherein the combination comprises (1) a population of donor cells modified to express a first isoform of a target protein (e.g., CXCR4) and (2) an antagonist (e.g., anti-CXCR4 antigen-binding protein) that specifically binds to a second isoform of the target protein but does not specifically bind to the first isoform of the target protein.

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.

This invention discloses a gene editing system for constructing porcine nuclear transplantation donor cells for cataract disease models with MIP gene mutations and its applications. This invention provides the application of MIP-gRNA2 (SEQ ID NO: 18), MIP-gRNA3 (SEQ ID NO: 19), and NCN protein in a preparation kit. This invention also provides a method for preparing recombinant cells, comprising the following steps: co-transfecting porcine cells with MIP-gRNA2, MIP-gRNA3, and NCN protein to obtain recombinant cells. The recombinant cells are recombinant cells with a mutated MIP gene. The kits described above are used for: preparing recombinant cells; preparing porcine cataract models; preparing cataract cell models, cataract tissue models, or cataract organ models. This invention has significant application value for the development of cataract drugs and for elucidating the pathogenesis of this disease.

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 technology includes a system to reallocate user experience resources. The system detects network performance in a set of cell areas. The system determines low-performing cell areas and high-performing cell areas by using a network performance model. The system determines common subscribers between the low-performing cell areas and the high-performing cell areas. Based on the common subscribers, the system identifies a recipient cell area and a donor cell area. The system determines a reallocation of user experience resources by using the network performance model to determine scores based on changes to cell site parameters of the donor cell area and the recipient cell area. The system generates an indication that the reallocation will improve user experiences for subscribers of the recipient cell area, without diminishing user experiences for subscribers of the donor cell area.