106 results about "Mesoderm" patented technology

The invention provides a construction method and application of a blood sinus networked liver organ. According to the method, pluripotent stem cells are co-differentiated into HAND1 < + > mesoderm and endoderm, a front intestine rear endoderm containing KDR < + > mesoderm progenitor cells is generated, a liver endoderm with CD34 < + > / CD31 < + > endothelial progenitor cells is generated, and the blood sinus networked liver organ is formed. The liver organ has a functional blood sinus endothelial network penetrating through the liver cluster, and has the characteristic functions of low-density lipoprotein uptake, human serum albumin removal, mediated secretion of blood coagulation factor 8 and the like. The method provides a new-generation highly bionic research tool for liver disease mechanism analysis, drug screening and regeneration repair.

The invention discloses a method for differentiating induced pluripotent stem cells into megakaryocytes and platelets, and relates to the technical field of induced pluripotent stem cell directional differentiation, and the method comprises the following steps: inoculating an iPSC single cell into a culture vessel, and carrying out adherent culture through a culture solution; after the cell adheres to the wall and before cloning is not formed, the liquid is changed into a first induction culture liquid, and the cell is induced and differentiated into a mesoderm cell; changing the solution into a second induction culture solution to induce and differentiate the mesoderm cells into hematopoietic endothelial cells; collecting the suspension cells, culturing the suspension cells in an amplification culture solution, and enriching megakaryocytes; inducing megakaryocyte polyploidy through a third induction culture solution; and culturing the polyploidy megakaryocyte through a fourth induction culture solution to promote platelet release. The method for inducing the pluripotent stem cells to differentiate into megakaryocytes and platelets is convenient and rapid, the megakaryocytes and platelets can be obtained through iPSC in a short time, and feeder cells, serum and additional gene editing are not needed; meanwhile, the yield is high.

The invention discloses a method for quickly and efficiently differentiating iPSC (induced pluripotent stem cells) into mesenchymal stem cells in a serum-free manner, the method comprises the following steps: firstly inducing the iPSC into a mesoderm precursor, then transferring the mesoderm precursor into a liquid culture medium for suspension culture, and finally obtaining iMSC (iPSC-doped MSC) by using an enrichment culture medium, and the method can shorten the differentiation time, improve the purity of the iMSC and obtain a large number of homogenized iMSCs.

The invention provides a cryopreservation and resuscitation method for induced differentiation of pluripotent stem cells to metaphase cells and application, and the cryopreservation method comprises the following steps: (1) the initial cells are pluripotent stem cells, and are induced and differentiated to the metaphase; (2) recovering the cells by using a mild digestive enzyme, counting, and resuspending the cells in a cryopreservation solution; and (3) carrying out programmed cooling to-80 DEG C, and then transferring into liquid nitrogen for preservation for later use. According to the cryopreservation method and the cryopreservation liquid provided by the invention, the cryopreservation survival rate of the differentiated metaphase cells is obviously improved, apoptosis or irreversible stress injury induced by a traditional cryopreservation liquid is avoided, and the problems of large cell injury and low survival rate caused by using a general cryopreservation liquid in the prior art are obviously improved. The resuscitated cells keep good differentiation potential, can smoothly form a mature renal unit structure, and solves the problem of subsequent differentiation failure in the prior art.

The invention relates to the technical field of stem cells, in particular to a CBA culture medium and application of the CBA culture medium in in-vitro induced differentiation of embryonic ectomesoderm cells. The invention provides a culture medium for rapidly (5 days) and efficiently (more than 90%) inducing the differentiation of the original state (Naive) and primed state (Primed) human pluripotent stem cells (hPSCs) into the extraembryonic mesoderm cells (ExMs) with self-renewal ability, the culture medium has universality, and simultaneously supports the induction of the rapid and efficient differentiation of the Naive and Primed hPSCs into the ExMs; the ExMs obtained by differentiating the hPSCs are highly similar to the ExMs in human embryos in the periimplantation stage and the periintestinal stage, and can be stably cultured and amplified for a long time in vitro. The culture medium and the induction method have the advantages of being short in period, high in yield and low in cost, can promote the industrialization of ExMs, and have wide application prospects.

The invention relates to the technical field of biology, in particular to a method for producing a platelet quality control product by using iPS cells and the platelet quality control product, and the method comprises the following steps: controlling overexpression of a specific gene in the iPS cells by using a gene editing technology to obtain an iPS cell line containing the specific iPS cells; a differentiation procedure is started by adding a culture medium containing a tetracycline derivative, and meanwhile, a fibroblast growth factor 2 and a bone morphogenetic protein 4 are added to promote mesoderm differentiation; growth factors and tetracycline derivatives are added to induce overexpression of specific gene segments and promote formation of specific megakaryocytes; adding an AhR antagonist and a ROCK inhibitor into the recovered specific megakaryocyte, and culturing to obtain platelets; and screening a platelet standard substance based on platelet functions and preparing the platelet standard substance into a quality control substance.

The invention belongs to the technical field of organoid culture, and particularly relates to a pulmonary alveolar organoid with an endogenous vascular network and a construction method of the pulmonary alveolar organoid. Aiming at the problems that mesoderm and endoderm are difficult to induce at the same time, the operation process is long, the difficulty is high and the like when a vascularized alveolar organ is constructed by an existing method, the invention provides the construction method of the alveolar organ with the endogenous vascular network. The method comprises the following steps: simultaneously and directionally inducing and differentiating human induced pluripotent stem cells (hiPSC) into fixed endoderm and mesoderm, anterior intestine endoderm, abdominal side anterior intestine endoderm and lung progenitor cells by adopting a one-step method, and promoting endothelial cell proliferation in an endogenous induced differentiation process through a specific growth factor combination. A functional vascular network is formed; and vascularized alveolar organs are induced to be generated. According to the invention, a qi-blood barrier structure in which vascular endothelial cells and pulmonary epithelial cells are tightly connected is successfully reproduced, the gas exchange function of lung tissues can be simulated, and a new way is provided for construction of vascularized alveolar organs.

Tissue specific progenitor cells generated from pluripotent sources representing endoderm, ectoderm and mesoderm lineages. Progenitor generation can be accomplished by contacting pluripotent stem cells with tissue-specific exosomes, microRNAs, proteins and peptides obtained from stressing said tissue ex vivo. Ex vivo generated tissue organoids which are utilized as sources of "differentiation factors" for the creation of personalized progenitor cells. For generation of pulmonary progenitor cells, decellularized cadaveric lung is populated with allogeneic stem cells to form pulmonary structures, said structures are exposed to various cellular stressors and conditioned media to differentiate pluripotent stem cells into progenitor cells. Extracellular matrix can be obtained from decellularized structures seeded with regenerative cells and used to create organoids that are subjected to stress in order to generate differentiating factors.

The application provides a freezing and recovery method and application of pluripotent stem cell induced differentiation middle stage cells, and the freezing method comprises the following steps: (1) the starting cells are pluripotent stem cells, which are induced to the middle stage; (2) the cells are recovered and counted by using a mild digestion enzyme, and the cells are resuspended in a freezing solution; (3) after programmed cooling to-80 DEG C, the cells are transferred into liquid nitrogen for storage, and are ready for use. The freezing method and the freezing solution provided by the application significantly improve the freezing survival rate of the differentiation middle stage cells, avoid the apoptosis or irreversible stress damage induced by the traditional freezing solution, and obviously improve the problems of cell damage and low survival rate caused by the general freezing solution in the prior art. The differentiation potential of the recovered cells is good, and the cells can successfully form mature kidney unit structures, so that the problem of subsequent differentiation failure in the prior art is solved.

Disclosed herein are improved in vitro methods of making splanchnic mesoderm cell types and subtypes thereof from pluripotent cells. These methods can be used to produced improved foregut- and hindgut-derived organoids containing enriched mesenchyme, which enhances organoid viability, growth, and maturation, both in in vitro culture and in vivo transplantation.

The invention provides a pluripotent stem cell embryoid differentiation method and a culture medium thereof, and belongs to the technical field of cell culture. The inhibitors (Chroman 1 and Emricasan) of two signal paths are added into the basic culture medium, so that the differentiation of the pluripotent stem cells to embryoid bodies (EB) can be promoted, the cells can keep a good state in the preparation and differentiation process, and the cell survival rate is improved. Compared with a traditional hanging drop method, a microwell plate method and a centrifugal method for preparing EB, the pluripotent stem cell embryoid body differentiation method provided by the invention is simpler and easy in process amplification, exogenous factors and serum are not involved in the whole process, target cells can be produced in a large scale while the clinical application safety is ensured, and the method is suitable for industrial production. And a good technical support is provided for clinical application of the pluripotent stem cell embryoid body and the germ layer cells thereof.

The invention provides a general induction method for original digestive tract partition. The method comprises the following steps: high-quality human pluripotent stem cell verification, pluripotent stem cell sensitivity test, cell inoculation and start convergence degree verification, endoderm-mesoderm co-induction, original digestive tract induction and original digestive tract partition. Compared with a traditional method, on the premise of not depending on serum, original digestive tract partition derivatives (in a 3D sphere form) of different organs such as lungs, stomachs, livers, pancreas, small intestines and large intestines can be efficiently generated, and subsequent batch preparation of final products is facilitated. The method is helpful for promoting large-scale industrial transformation and application of respiratory digestive tract organs.

The invention discloses a method for inducing iPSCs (induced pluripotent stem cells) to directionally differentiate into vascular endothelial cells by using a small molecule compound and cell factor composition, which is characterized in that iPSCs cells are used as starting cells, and a vascular endothelial cell differentiation scheme completely dependent on chemical small molecule and cell factor stimulation is established. An E6 culture medium containing CHIR-99021, BMP4 and Activin A is utilized to induce mesoderm progenitor cells, an E6 culture medium containing VEGFA (vascular endothelial growth factor A), FGF-2, 8-Bromoadenosine 3 ', 5'-cyclic monophosphate monophosphate, SB431542, Forskolin and Melatonin is utilized to induce the endothelial cells, and an E6 culture medium containing VEGFA, FGF-2 and Hydrocortisone is utilized to further induce and differentiate the endothelial cells into the vascular endothelial cells. By using the scheme, the differentiation efficiency can exceed 99%, and differentiated cells have phenotypes and functions of vascular endothelial cells and do not express pluripotent related markers SSEA-4, TRA-1-60 and TRA-1-81.

This invention discloses a method for inducing pluripotent stem cells to differentiate into skin keratinocytes and its culture medium, solving problems such as long differentiation cycles, low yields, insufficient purity, and animal-derived contamination in existing technologies. The first stage uses medium I containing FGF2 and tanshinone IIA to promote mesodermal differentiation. The second stage uses medium II containing BMP4, EGF, dexamethasone, and PVA to regulate and synergistically induce the BMP / ERK signaling pathway. The final stage uses medium III for expansion culture, obtaining fibroblasts with a purity >99% within 20 days. This method is non-invasive, simple, and low-cost, with potential for large-scale production. The obtained cells can be used for skin regeneration, disease modeling, drug screening, and clinical treatment, demonstrating significant medical application and industrial transformation value.

The invention provides a 3D culture method for promoting stem cells to be differentiated into cardiac lineage cells. According to the method, four-stage time sequence induction is adopted, wherein in the stage I, BMP4, Activin A and bFGF are used for efficiently inducing the cells into mesoderm in an insulin-free culture medium; in the stage II, DKK1 is used for inhibiting WNT signals to lock the heart pedigree, and VEGF165 is added in the early stage to start cardiac endothelial differentiation; in the stage III, metabolic support is provided for an insulin-containing culture medium, and bilineage maturation is deepened; in the IV stage, bFGF is added to support long-term survival and function maintenance of the cells, and the whole process is carried out in a 3D embryoid body. According to the invention, the cheap RPMI1640 basal culture medium is used for replacing an expensive commercial culture medium, the high-purity heart micro-tissue which can spontaneously beat and contains cardiac muscle and cardiac endothelial cells can be obtained within about 10 days, and an economical and efficient cell source is provided for heart disease research, drug screening and regenerative medicine.

The invention relates to the field of biomedicine, in particular to a construction method of an in-vitro embryo model. The invention provides a construction method of an in-vitro embryo model. According to the construction method, nerve-induced cells and tail-side mesoderm-induced cells are co-cultured under specific conditions to obtain the in-vitro embryo model. The method has relatively high expansibility, high efficiency and repeatability, so that gene modification aiming at a specific pedigree becomes possible, and the method has huge application potential in the field of regenerative medicine.

The present application relates to the technical field of induced pluripotent stem cell, and particularly relates to mesoderm differentiation specific human induced pluripotent stem cell and application thereof. The present application provides human induced pluripotent stem cell BC-hiPSC-ME-20, which is preserved in China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.46536. The cell strain has multi-lineage differentiation potential and high-efficiency mesoderm directional differentiation capacity, has stable pluripotency and excellent passage stability, and has higher safety, and has a good application prospect in disease model construction, disease occurrence and development mechanism research, drug screening and cell therapy.

Tissue-specific progenitor cells representing the endoderm, ectoderm, and mesoderm lineages are generated from pluripotent sources. Progenitor cell generation is accomplished by contacting pluripotent stem cells with tissue-specific exosomes, microRNAs, proteins, and peptides, obtained by stressing the tissue under in vitro conditions. In vitro-generated tissue organoids are used as a source of "differentiation factors" for generating individualized progenitor cells. For example, to generate lung progenitor cells, allogeneic stem cells are implanted into decellularized cadaveric lung tissue to form lung structures, which are then exposed to various cellular stressors and conditioned media to differentiate the pluripotent stem cells into progenitor cells. Extracellular matrix can be obtained from decellularized structures seeded with regenerating cells and used to generate organoids, which are then subjected to stress to produce differentiation factors.

The present disclosure relates to in vitro co-culture models and methods for generating such co-culture models. Co-culture models of this disclosure, and related methods, may include co- culturing or maintaining different cell types in a culture receptacle in a common culture medium. In a specific example, a co-culture model (and related methods) may include co- culturing or maintaining one, two, or more mesoderm lineage cell-types and at least one endoderm lineage cell-type in a culture receptacle in a common culture medium.

The present disclosure provides biomaterials and methods for preventing and minimizing progression of cartilage and / or connective tissue damage. Also provided herein are biomaterials and methods for alleviating and / or reducing the risk for developing arthritis (e.g., osteoarthritis) associated with joint injury and / or joint surgery.

The application belongs to the field of stem cell biology and relates to a culture method of a 3D nerve immune organoid containing microglia cells, which comprises the following steps: S1, inducing induced pluripotent stem cells in ectoderm direction and mesoderm direction respectively to obtain nerve-like embryoid bodies and yolk sac-like embryoid bodies; S2, culturing the nerve-like embryoid bodies to make them differentiate into nerves to obtain nerve ring structures, re-digesting the nerve ring structures to obtain nerve progenitor cell single cells, and culturing the yolk sac-like embryoid bodies to make them generate macrophage progenitor cells; and S3, fusing the nerve progenitor cell single cells and the macrophage progenitor cells and continuing to culture to form the 3D nerve immune organoid containing microglia cells. The brain-like organoid of the application can realize the following functions: exploring the control of microglia cells on the proportion of progenitor cells in the development stage, the change of phagocytosis, the control on the number of mature neurons, the influence on cell proliferation and apoptosis, and simultaneously accepting external stimulation and making corresponding functional stress changes.

The invention relates to the technical field of biology, in particular to a method for modular assembly of artery blood vessel type organs and construction of tissue microcirculation of the artery blood vessel type organs. The method comprises the following steps: inducing human pluripotent stem cells to differentiate through a mesoderm stage to form an arterial specific vascular organoid module (AVO), and then carrying out modular assembly on the AVO in vivo or in vitro so as to quickly reconstruct a tissue microcirculation network with arterial characteristics. The obtained artery blood vessel organoid is uniform in form, the diameter of the artery blood vessel organoid is intensively distributed at about 200 microns, SOX17 and CXCR4 artery endothelial markers are expressed at the same time, and the artery blood vessel organoid has polarity and a complete basilar membrane. Transplanting experiments show that the organ can be integrated with host blood vessels in a short time, and ischemic tissue blood perfusion is recovered. The defects that existing organoid is irregular, difficult to splice, delayed in perfusion and the like are overcome, and a new strategy is provided for revascularization and acute ischemic disease repair.

The invention relates to the technical field of plant genetic engineering, in particular to application of a plant HSF family transcription factor HSFA2B in regulation and control of temperature response growth of plant mesocotyl. The invention provides application of a plant HSF family transcription factor HSFA2B in regulating and controlling temperature response growth of mesocotyl in a plant seedling stage or regulating and controlling direct seeding emergence of plants at high environment temperature. The invention finds that HSFA2B has an important function in rice mesocotyl response temperature growth regulation and control. The mesocotyl of the mutant with the HSFA2B knocked out is obviously longer than that of the wild type at the proper environment temperature, but the mesocotyl of the mutant with the HSFA2B knocked out is not obviously different from that of the wild type at the high environment temperature, and the response sensitivity to the high environment temperature is reduced. The discovery of the new function of the HSFA2B is of great significance to the breeding and direct seeding production of rice direct seeding varieties.

This invention discloses a method for constructing lung organoids with spontaneous vascularization and branching morphology. By optimizing the culture protocol and progressively inducing directional mesoderm, vascularized foregut endoderm, vascularized lung bud organoids, and vascularized lung organoids, lung organoids with both vascularization and branching morphology are successfully cultured, thus more realistically simulating the structural characteristics of the human lung. This invention has significant advantages: First, it can effectively induce the formation of vascularization and branching structures in lung organoids, overcoming the shortcomings of traditional lung organoids that lack vascularization and branching structures; second, by adjusting the culture method in the early stages, the uniformity of the lung organoids is significantly improved, enhancing the reliability and reproducibility of experimental results. This invention provides a more precise and efficient experimental platform for preclinical studies such as lung disease research, drug screening, and toxicology testing, and has significant application value.

The disclosure relates to a cell culture method for deriving a multipotent neuromesodermal progenitor (NMP) cell and / or lineages of specific anterior-posterior positions along the human body axis and / or cell types of matched anterior-posterior position, from a human pluripotent stem cell (PSC), comprising culturing said stem cell under conditions and with reagents modulating the following signalling pathways: (i) activation of FGF signalling for a duration of about 12-120 hours; (ii) activation of WNT signalling for a duration of about 48-120 hours; (iii) inhibition of SMAD signalling, including TGF-beta / activin-Nodal signalling and BMP signalling, for a duration of about 48-120 hours, wherein said progenitor co-expresses SOX2, TBXT, CDX2 and CDX1 at RNA and / or protein level(s).The disclosure further relates to a composition for use in deriving multipotent NMPs and / or daughter lineages of a specific anterior-posterior position in the human body and / or cell types of matched anterior-posterior position from human pluripotent stem cells (PSC), a multipotent NMP, a cervical and a thoracic NMP obtained by culturing human PSCs, a composition for use in generating lumbar level NMPs from the multipotent NMPs, a lumbar level NMP obtained by culturing the multipotent NMP, a composition for use in generating tbx6 expressing presomitic mesoderm cells from the multipotent NMPs, a tbx6 expressing presomitic mesoderm cell obtained by culturing the multipotent NMP, a human cell derived from the multipotent NMP, a cell population, a kit and a cellular composition.

The invention discloses a vascular endothelial cell as well as a preparation method and application thereof. The preparation method comprises the following steps: inoculating induced pluripotent stem cells into a culture medium containing an ROCK signal channel inhibitor for culturing, and inducing the cultured induced pluripotent stem cells by using a culture medium containing a Wnt signal channel activator and a BMP signal channel activator, so that the induced pluripotent stem cells are differentiated into mesoderm cells; inducing the mesoderm cells by using a culture medium containing a vascular endothelial growth factor and a cAMP signaling pathway agonist, so that the mesoderm cells are differentiated into endothelial coarse cells; and performing positive sorting of the endothelial cell marker CD144 on the endothelial coarse cells to obtain purified vascular endothelial cells. The induction path of the method is clear and efficient, the period is short, the vascular endothelial cell population with high purity, complete vascular barrier function and mature phenotype can be obtained, and the cell population can be applied to construction of a vascular network in a gel matrix.

The application discloses a lipoic acid injection composition applied to mesoderm and a preparation method thereof, wherein the lipoic acid injection composition comprises lipoic acid, cyclodextrin, a stabilizer, a regulator, a pH regulator and an osmotic pressure regulator. The lipoic acid powder is dissolved by using the cyclodextrin, the solubility of the lipoic acid is improved, the post-stability of the injection is improved by using the wrapping property of the cyclodextrin, meanwhile, the irritation of the cyclodextrin to the mesoderm is reduced by adding hyaluronic acid and a salt thereof, and the comfortable feeling of a user is obviously improved.

This document relates to methods and materials for making and using cardiopoetic stem cells. For example, methods and materials for delivering one or more nucleic acids (e.g., one or more RNAs) encoding one or more early mesodermal transcription factors to a stem cell to generate a cardiopoetic stem cell are provided.

The invention relates to the technical field of human induced pluripotent stem cells, and discloses a method and a system for constructing vascular organs based on human induced pluripotent stem cells, and the method comprises the following steps: inducing human induced pluripotent stem cells to aggregate into cell aggregates, and identifying aggregate characteristics of the cell aggregates; determining mesoderm induction molecules of the cell aggregate to construct a mesoderm induction environment of the cell aggregate, and performing induced differentiation on the human induced pluripotent stem cells in the cell aggregate; detecting the expression level of the mesoderm marker gene in the mesoderm cells, constructing a vascular induction culture medium of the mesoderm cells, and inducing the mesoderm cells to differentiate into vascular lineage cells; constructing a three-dimensional extracellular matrix of the vascular lineage cells to differentiate the vascular lineage cells into a vascular network; and calculating the permeability and branch density of the vascular network to determine the fluid shear force and pulse frequency of the vascular network, and culturing the vascular network into a vascular organ. The construction stability and functionality of the vascular organoid can be improved.

The invention relates to the technical field of induced pluripotent stem cells, in particular to a mesoderm differentiation specific human induced pluripotent stem cell and application thereof. The invention provides a human induced pluripotent stem cell BC-hiPSC-ME-20, which is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.46536. The invention further provides a preparation method of the human induced pluripotent stem cell BC-hiPSC-ME-20. The cell strain has multilineage differentiation potential and efficient mesoderm directional differentiation capacity, has stable pluripotency and excellent passage stability, has high safety, and has good application prospects in disease model construction, disease occurrence and development mechanism research, drug screening and cell therapy.