421results about "Pancreatic cells" patented technology

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 invention belongs to the technical field of organoid culture, and particularly relates to a co-differentiated vascularized islet organoid and a construction method thereof. Aiming at the problems that the existing pancreatic islet organ vascularization mostly adopts a co-culture mode, the operation process is long, the difficulty is high, the randomness is realized, the uniformity is poor and the like, the invention provides the construction method of the co-differentiated vascularized pancreatic islet organ. The method comprises the following steps of: sequentially inducing and culturing directional endoderm cells, original intestinal canal cells and posterior preintestinal cells, and adding a culture medium containing VEGF (vascular endothelial growth factor), BMP4 (bone morphogenetic protein 4) and FGF2 (fibroblast growth factor 2) for culturing in subsequent induced differentiation, so as to obtain the co-differentiated vascularized islet organ. Under the condition that exogenous endothelial cells are not added, the iPS cells are directionally differentiated to generate the vascularized islet organ containing beta, alpha and delta cells and vascular endothelial cells for the first time, the vascularized islet organ is more mature than non-vascularized islet organs and has a stronger insulin secretion function, and a new thought and an experimental basis are provided for realizing functional healing of diabetes mellitus.

According to the sgRNA highly targeting the IFNGR1 gene, the composition and the application of the sgRNA, the sgRNA provided by the invention can highly target the IFNGR1 gene, complete knockout of IFNGR1 on mixed cloning is realized through combined application of three sgRNAs, and the process and the time of constructing an IFNGR1 knockout cell line are greatly shortened and shortened.

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.

Disclosed herein are compositions and methods related to differentiation of stem cells into pancreatic islet cells. In some aspects, the methods provided herein relate to generation of pancreatic β cell, α cell, δ cells, and EC cells in vitro. In some aspects, the disclosure provides pharmaceutical compositions including the cells generated according to the methods disclosed herein, as well as methods of treatment making use thereof.

The invention discloses a double-chamber micro-fluidic organ chip, a preparation method and application, and belongs to the technical field of biological materials. The double-chamber micro-fluidic organ chip sequentially comprises a top layer, an upper layer, a middle layer and a lower layer from top to bottom, cavities are formed in the upper layer and the lower layer, and micropore arrays for accommodating biological samples are arranged in the cavities; the chip further comprises a plurality of circulation channels which can be respectively communicated with the upper layer chamber and the lower layer chamber, so that a biological sample enters and exits from the micropore array; and the middle layer is a semi-permeable porous membrane. The micro-fluidic organ chip can be used for establishing a metabolic disease model, promoting interaction between organs and realizing space-time dynamic change analysis of tissues and organs.

Disclosed herein are compositions and methods related to differentiation of stem cells into pancreatic islet cells. In some embodiments, the methods provided herein relate to generation of pancreatic β cell, α cell, δ cells, and EC cells in vitro. In some embodiments, the disclosure provides pharmaceutical compositions including the cells generated according to the methods disclosed herein, as well as methods of treatment making use thereof.

The invention relates to the field of biology, and provides a human immortalized pancreatic cancer fibroblast as well as a preparation method and application thereof.The human immortalized pancreatic cancer fibroblast comprises three cell strains which are all primary fibroblast cells extracted from tumor tissues of pancreatic ductal adenocarcinoma patients, the human pancreatic cancer fibroblast cell strain CAFPC-2209131, the human pancreatic cancer fibroblast cell strain CAFPC-2209132 and the human pancreatic cancer fibroblast cell strain CAFPC-2303221 are classified and named as the human pancreatic cancer fibroblast cell strain CAFPC-2209131, the human pancreatic cancer fibroblast cell strain CAFPC-2209132 and the human pancreatic cancer fibroblast cell strain CAFPC-2303221 respectively, the human pancreatic cancer fibroblast cell strain CAFPC-2209132 and the human pancreatic cancer fibroblast cell strain CAFPC-2303221 are all Cell STR identification and immunofluorescence detection prove that the cell strain is a fibroblast cell strain. After the tumor-associated fibroblast and pancreatic cancer cells are co-cultured, the proliferation ability of the tumor cells is obviously enhanced, and the sensitivity of the tumor cells to a chemotherapeutic drug gemcitabine is reduced. A new thought is provided for diagnosis and treatment of pancreatic cancer, and based on the pancreatic cancer in-vitro co-culture model established in vitro, the application has important significance for researching the effect and related mechanisms of fibroblasts in the pancreatic cancer tumor microenvironment.

The invention belongs to the technical field of biomedicine, and relates to a method for increasing the yield of pancreas islet organs differentiated by pluripotent stem cells and improving the stability. The method comprises the following steps: (1) inoculating pluripotent stem cells, and culturing by adopting a basic culture medium containing Y-27632; (2) changing the liquid, and continuously culturing by adopting a basic culture medium; (3) culturing for 1 day by using an S1D1 culture medium, and then culturing for 2-3 days by using an S1D2 culture medium; (4) changing the liquid, and culturing for 2-3 days by adopting the S2 culture medium; (5) changing the liquid, and culturing for 1-2 days by adopting an S3 culture medium; (6) changing the liquid, and culturing for 4-7 days by adopting a culture medium S4; (7) changing the liquid, and culturing for 3-10 days by adopting an S5 culture medium; and (8) carrying out cell sphere repolymerization, and culturing for 3-21 days by using an S6 culture medium to obtain the pancreas islet organ. The present invention increases the yield and differentiation efficiency at each differentiation stage and increases the scale yield at the cellular level.

The present disclosure provides methods of producing a population of endodermal stem cells (EnSC), a population of EnSC-derived PP cells, a population of EnSC-derived EP cells, or regenerative pancreatic islet tissue (E-islets). The disclosure also relates to EnSC, EnSC-derived PP cells, EnSC-derived EP cells or regenerated pancreatic islet tissue obtained using the method, and therapeutic uses thereof.

The invention relates to a polypeptide derivative for pancreas islet surface modification and inhibition of pancreas islet transplantation IBMIR reaction, and belongs to the technical field of biological medicine. The peptide derivative disclosed by the invention is obtained by carrying out an amide reaction on KF7 peptide and levodopa, and the amino acid sequence of the KF7 peptide is shown as SEQ ID NO: 1. The polypeptide derivative has good biocompatibility, the surface modification method is simple, the modification process has no influence on cell life activity, activation, aggregation, expansion and adhesion of platelets can be inhibited, the bleeding risk is reduced, a blood coagulation system is not influenced, pancreas islet intrahepatic colonization and pancreas islet functions after portal transplantation are enhanced, and the application prospect is wide. The composition can protect pancreas islet, prevent infiltration of platelets and CD11b + cells, inhibit pancreas islet transplantation IBMIR reaction, control blood sugar level, and prevent and treat diabetes mellitus.

The present disclosure provides modified cells comprising a nucleic acid encoding an agent (e.g., a detection agent, a selection agent, or a detection agent and a selection agent) inserted at an endogenous proliferation gene or off-target cell marker gene, as well as compositions, methods, uses, and kits related thereto.

The present disclosure provides methods of making tissue-derived epithelial organoids. In particular, the present disclosure provides a tissue-derived epithelial organoid embedded in a hydrogel suspended in a culture medium. The present disclosure further provides methods of using such organoids.

The invention relates to the field of cell biology, and particularly relates to a preparation method of a pancreatic cancer-related antigen-loaded polypeptide, which is characterized in that factors such as GM-CSF, IL-4, IFN-gamma, TNF-alpha, IL-1beta, IL-6, PGE2, Poly I: C, R848 and the like are added to promote mononuclear cells to be activated into DC cells, pancreatic cancer PANC-1, MIA PaCa-2, BxPC-3, KP4 and SW1990HM antigens are loaded, and not only is DC cell maturation promoted, but also pancreatic cancer-related antigens are loaded. The positive rate of CD83 + CD86 of the DC cells cultured by the method is 94.16%, and the secretion amount of IL-12p70 in a cell supernatant is (645.38 + / -36.49) pg / mL. By adopting the culture method disclosed by the invention, the maturation rate of the DC cells can be improved while the DC cells are activated, so that the culture method for promoting the maturation and amplification of the DC cells loaded with the pancreatic cancer antigen is achieved.

Disclosed herein are intravascular artificial pancreas devices and methods for their manufacture.

Embodiments herein describe tools, instruments and methods for aseptic loading, dispensing and / or delivering cells into an implantable device and aseptically and selectively scaling a device inside a sterile package as well as and storing and preparing for shipment the cell-filled device.

The present disclosure relates to a transplantable cell therapy product composition for diabetes mellitus that contains an aggregate of insulin-secreting cells derived from stem cells. In the present disclosure, an NF microwell array membrane was fabricated by applying a molding process to an electrospun, permeable, biodegradable polycaprolactone (PCL) NF membrane and thus allows gases and soluble factors to permeate therethrough. The NF microwell of the present disclosure could provide more nutrients to the iPSC aggregates than conventional impermeable PDMS microwells, thus enhancing survival and differentiation capabilities of the cells. Additionally, the NF membrane was attached singly to the subcutaneous tissue and to the surface of organs such as liver and peritoneum without the need for a fixing material or separate sutures and was integrated with surrounding tissues, resulting in higher insulin secretion than PDMS microwells. Therefore, the present disclosure can be effectively utilized as a composition for the prevention or treatment of diabetes.

Disclosed herein include methods, compositions, and kits suitable for use in cell therapy. In some embodiments, there are provided methods and compositions for differentiating stem cells into pancreatic beta cells capable of producing insulin.

The present disclosure provides methods of amplifying tissue samples derived from humans for purposes of treating certain diseases. The method comprises: isolating a small amount of associated tissue by biopsy or other tissue acquisition methods; the tissue is amplified in vitro, and meanwhile, natural functions are reserved; and re-implanting the amplified tissue to alleviate human disease. One example includes ex vivo expansion of human pancreatic islet tissue and reimplantation of the expanded cells into an individual with Type 1 diabetes for treatment and long term relief or cure of disease.

The invention provides for methods of differentiating pancreatic endocrine cells into pancreatic beta cells expressing PDX1, NKX6.1, MAFA, UCN3 and SLC2A. These pancreatic beta cells may be obtained by step-wise differentiation of pluripotent stem cells. The pancreatic beta cells exhibit glucose-dependent mitochondrial respiration and glucose-stimulated insulin secretion similar to islet cells.

The application provides an islet-vascular co-culture system based on a microfluidic chip, a building method and application, and the building method of the islet-vascular co-culture system comprises the following steps: S1, preparing microfibers loaded with islets; S2, preparing an assembled chip, the assembled chip comprises a containing cavity, first microchannel networks and second microchannel networks are symmetrically arranged on two sides of the containing cavity, and the first microchannel networks and the second microchannel networks are respectively communicated with a liquid inlet and a liquid outlet at an end away from the containing cavity; S3, controlling the flow of the liquid inlet to be 6-8 muL / min to carry out dynamic culture. The application can simulate the key structure and characteristics of islet tissue, can evaluate a diabetes treatment drug in vitro, can bridge the gap between in vitro and in vivo models, and can provide new tools and methods for the fields of drug development, disease model construction and personalized medical treatment.

The invention provides a microfiber for loading pancreas islet, a preparation method and application, the preparation method comprises the following steps: S1, preparing a micro-fluidic chip and building a pneumatic control micro-fluidic spinning device, the micro-fluidic chip is provided with channels for conveying a core flow, a sample flow and a sheath flow, the micro-fluidic chip further comprises a PDMS chamber, and the PDMS chamber is provided with a PDMS cavity; a channel for periodically pressurizing to expand so as to squeeze and close the core flow; s2, adding collagen into the core flow, then adding beta-TC6 cells to enable the cell density of the core flow to be 4 * 10 < 6 > mL <-1 > to 8 * 10 < 6 > mL <-1 >, respectively controlling the flow rates of the core flow, the sample flow and the sheath flow to be 0.8-1.2 [mu] L / min, 18-22 [mu] L / min and 80-120 [mu] L / min, and carrying out preparation under the frequency of 0.8-1.2 Hz; according to the method, a pneumatic control droplet generation method is combined with microfluidics, so that the high-flux islet-like can be prepared more simply and quickly, and the difference between the islet-like and an in-vivo complex microenvironment is small; the prepared microfiber is continuous and complete, and a large number of water-phase liquid drops containing beta-TC6 cells are uniformly distributed in the microfiber.

Disclosed herein are compositions and methods related to differentiation of stem cells into pancreatic endocrine cells. In some aspects, the methods provided herein relate to generation of pancreatic β cell, α cell, δ cells, and EC cells in vitro. In some aspects, the disclosure provides pharmaceutical compositions including the cells generated according to the methods disclosed herein, as well as methods of treatment making use thereof.

The present disclosure relates to an in vitro method for enhancing engraftment of isolated pancreatic cells comprising the step of contacting an isolated pancreatic cell prior to a transplantation in a subject in need thereof, with a gem-difluorinated C-glycopeptide compound of general formula I, or a pharmaceutically acceptable base, addition salt with an acid, hydrate or solvate of the compound of general formula I:

The present invention provides methods to promote the differentiation of pluripotent stem cells. In particular, the present invention provides methods to produce a population of cells, wherein greater than 10% of the cells in the population express markers characteristic of single hormonal pancreatic beta cells.

The present invention provides a method for lowering blood glucose levels in an animal by transplanting a population of pancreatic endocrine precursor cells into an animal.

The invention provides for methods of differentiating pancreatic endocrine cells into pancreatic beta cells expressing PDX1, NKX6.1, MAFA, UCN3 and SLC2A. These pancreatic beta cells may be obtained by step-wise differentiation of pluripotent stem cells. The pancreatic beta cells exhibit glucose-dependent mitochondrial respiration and glucose-stimulated insulin secretion similar to islet cells.

A novel method of inducing or producing hepatocytes from human induced pluripotent stem cells at an unprecedented efficiency and functionality. The core of the invention is the use of experimentally discovered mRNAs at multiple critical differentiation decision points along a pluripotent to mesendoderm to endoderm to hepatocytes pathway in a previously unknown manner.

The application provides a preparation method of organoids based on a single cell chip, which comprises microfluidic chip preparation, a two-water-phase solution preparation, a single cell suspension preparation, microfluidic manipulation, single cell-loaded microgel generation and organoid construction and the like. The application takes a microfluidic chip integrated with a normally closed pneumatic pump valve as a technical platform, takes a single cell suspension pre-incubated with a crosslinking agent as a chemical reaction core, takes a water-in-water droplet as a forming template, and realizes efficient in-situ loading of single dispersed seed cells in a micro hydrogel carrier through a one-step method, and simply removes empty droplets without cells in a subsequent transfer step. The single cells in the microgel can be cultured, expanded and self-organized to form required organoids. The application improves the consistency of the starting point of the organoids, has the high-throughput characteristics of the droplet microfluidic technology, realizes efficient and highly reproducible in-vitro construction of the organoids, and can play a great role in the fields of basic research and transformation application related to the organoids.