76 results about "Mesoderm" patented technology

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 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 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 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.

This invention discloses a fungal-derived antibacterial blue fluorescent carbon dot, its preparation method, and its application. The carbon dot is synthesized by heating an extract of *Metacarpus mesoderm*. Specifically, this invention uses a natural product extract of the endophytic bacterium YSP3 as a raw material and synthesizes blue fluorescent carbon dots (Du-CDs) using a one-step solvothermal method. The average particle size of these Du-CDs is 13.9 nm, and they are positively charged. The Du-CDs prepared by this invention can efficiently kill bacteria and fungi, exhibiting broad-spectrum antibacterial properties. This invention provides carbon dots with better bactericidal effects and wound-healing capabilities compared to natural fungal extracts, along with good biocompatibility, and develops novel antibacterial drugs with good fluorescence.

This invention provides a universal method for inducing primitive digestive tract compartments. The method comprises the following steps: high-quality human pluripotent stem cell assay, pluripotent stem cell sensitivity testing, cell seeding and initial confluence testing, endomesoderm co-induction, primitive digestive tract induction, and primitive digestive tract compartmentalization. Compared to traditional methods, this method, without relying on serum, can efficiently generate primitive digestive tract compartmentalized derivatives (in the form of 3D spheroids) of various organoids, such as the lung, stomach, liver, pancreas, small intestine, and large intestine, facilitating subsequent mass production of the final product. This invention will help promote the large-scale industrial application and application of aerodigestive tract organoids.

The present disclosure relates to a method for induced differentiation of stem cells into hematopoietic progenitor cells. The method comprises the following steps: culturing pluripotent stem cells to obtain an embryoid body; performing mesoderm differentiation culture on the embryoid body to obtain mesoderm cells; performing hemogenic endothelium differentiation culture on the mesoderm cells to obtain hemogenic endothelial cells; and performing hematopoietic progenitor cell differentiation culture on the hemogenic endothelial cells to obtain hematopoietic progenitor cells. According to the present disclosure, the hematopoietic progenitor cells can be rapidly and efficiently prepared, and the prepared hematopoietic progenitor cells have an ability to stably differentiate into a plurality of different blood cells (including simultaneously having erythroid, myeloid, and lymphoid cells); by optimizing a culture system, the differentiation efficiency is significantly improved and the number of hematopoietic progenitor cells obtained is significantly increased.

The invention relates to mesodermal killer (MK) cells and their use in therapy, especially for the treatment of cancer.

The invention discloses a bionic intervertebral disc integrated organoid based on 3D printing and a construction method and application thereof, and relates to the field of biomedical engineering. The construction method comprises the following steps that a polymer material is adopted to prepare a crossed net-shaped bionic stent through 3D printing, the crossed net-shaped bionic stent is curled to form a cylindrical fiber ring structure, and the fiber ring bionic stent is obtained; performing mesoderm induction on the pluripotent stem cells, and respectively performing cartilage induction and fibrous cartilage induction to obtain nucleus pulposus-like cartilage organs and fibrous ring-like fibrous cartilage organs; the cartilage-like organ is placed in the center of the fibrous ring bionic scaffold, the fibrous cartilage-like organ is filled in the grid structure of the fibrous ring bionic scaffold, and after matrigel wrapping and incubation curing, the bionic intervertebral disc integrated organ based on 3D printing is obtained through induction culture. The bionic intervertebral disc integrated organ based on 3D printing provided by the invention can effectively reproduce structural characteristics and functional characteristics of a natural intervertebral disc.

A method for mesenchymal stem cell differentiation from human pluripotent stem cells is provided, particularly including the step of adaptation of embryoid bodies from 3-dimensional culture to 2-dimensional culture. The method enables rapid mesoderm and mesenchymal stem cell transformation by transferring from 3-dimensional culture to 2-dimensional culture. Also, the method relates to obtaining mesenchymal stem cell, which can be used in clinical applications, with this method.

The present invention provides a method for constructing a sinusoidal network liver organoid and its application, wherein pluripotent stem cells are co-differentiated into HAND1 + Mesoderm and endoderm produce KDR + Mesodermal progenitors give rise to the posterior foregut endoderm, which has CD34 + / CD31 + Endothelial progenitor cells form hepatic endoderm and form a sinusoidal network in liver organoids. These liver organoids possess a functional sinusoidal endothelial network throughout the liver clusters, exhibiting characteristic functions such as low-density lipoprotein uptake, human serum albumin clearance, and secretion of coagulation factor 8. This method provides a new generation of highly biomimetic research tools for understanding liver disease mechanisms, drug screening, and regenerative repair.

The invention discloses a preparation method and application of renal progenitor cells. The method specifically comprises the following steps: inducing pluripotent stem cells to be differentiated into an intermediate mesoderm cell stage through a GSK-3alpha / beta inhibitor, and inducing cells in the intermediate mesoderm cell stage to be differentiated into a cap-like mesenchymal stage by using a nuclear receptor agonist containing FGF-2 and RAR / RXR, so as to obtain the renal progenitor cells with obvious specific characteristics and potentials of the cap-like mesenchymal. According to the method, the high-purity and high-quality renal progenitor cells can be obtained without separation and purification, the purity can reach 99.9%, and the obtained renal progenitor cells have the differentiation potential of kidney-like organs, basically have no PSCs residues, have the differentiation potential of kidney-like organs and are suitable for clinical cell therapy; and the operation is simple, stable and controllable, and safe, effective, high-purity and homogeneous renal progenitor cells can be prepared on a large scale.