48 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 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.

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.

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 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 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 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 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.

Provided herein are compositions of anorectal transition zone stem cells, to treat endodermal, ectodermal or mesodermal disease. Also provided are cellular compositions of the anorectal transition zone stem cells, in a pharmaceutically acceptable carrier.

The present invention relates to: a method for differentiating stem cell-derived hepatocytes by using fasudil and gelatin; and the like. The method for differentiating stem cell-derived hepatocytes, according to the present invention, induces, by means of fasudil, the differentiation of stem cells into endoderm in an early stage of differentiation of stem cells into hepatocytes, inhibits lipid droplet formation through high-viscosity gelatin coating of a culture container, activates mitochondrial functions, up-regulates the expression of endoderm genes, and down-regulates the expression of ectoderm and mesoderm genes, thereby improving the efficiency of differentiation of stem cells into hepatoblasts. In addition, low-viscosity gelatin is added to the medium in a differentiation and maturation (late) stage so as to suppress the accumulation of lipid droplets, mitochondria are activated to improve the efficiency of differentiation of hepatoblasts into hepatocytes, and the function of differentiated hepatocytes is enhanced to improve the efficiency of differentiation of stem cells into hepatocytes. Therefore, hepatocytes obtained using the method are expected to be effectively used as a cell therapeutic agent, a transplant, and the like for treating liver diseases.

This document relates to methods and materials for making and using cardiopoietic 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 cardiopoietic stem cell are provided.

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 obtaining, activating and amplifying natural killer cells through iPSC differentiation induction, and relates to the technical field of stem cell biology, the method for obtaining, activating and amplifying natural killer cells through iPSC differentiation induction comprises the following steps: S1, preparing and inoculating iPSC cells; s2, differentiating the iPSC to a mesoderm; s3, the mesoderm cells are differentiated to CD34 + hematopoietic endothelial cells; s4, differentiating the CD43 < + > / CD45 < + > hematopoietic stem cells into the iNK cells; s5, maturation of the iNK cells; s6, activating and amplifying the iNK cells; in the step S1, cells are resuspended by using a fresh E8 complete culture medium and are inoculated into a porous culture plate; a plurality of induction culture media are used in the culture process; by adopting a porous culture plate and using an E8 complete culture medium to resuspend cells, efficient inoculation and primary culture of iPSC are realized, and the survival rate and amplification efficiency of cells are improved; the differentiation process of the iPSC towards the mesoderm, the hematopoietic endothelial cells and the CD34 + hematopoietic stem cells is optimized by using various induction culture media, and the differentiation efficiency is improved.

The present invention relates to the field of molecular biology, and in particular to a small-molecule combination, kit and method for directed differentiation of lymphohematopoietic precursor cells. A culture system comprises a sphere-forming medium, a mesoderm induction medium, a hematopoietic endothelial cell induction medium, a hematopoietic stem / progenitor cell induction medium, and a lymphohematopoietic precursor cell induction medium. In the present invention, by modulating the composition of culture medium and the formulation of growth factors, cells can obtain sufficient nutrients and signals during induction, thereby improving the differentiation efficiency and stability. Using full suspension culture to simulate a three-dimensional in vivo environment avoids the problems of cell morphology change and stress while enhancing the stability and growth rate of cells.

1. The name of the design product: water light dot matrix instrument (BTS). 2. The use of the design product: for mesoderm care of skin. 3. The design points of the design product: in shape. 4. The picture or photo that best shows the design points: perspective view 1.

The invention provides a preparation method of macrophages derived from pluripotent stem cells. Dynamic culture is combined in the process that an embryoid body is sequentially differentiated into mesoderm progenitor cells, hematopoietic stem cells and myeloid cells, a specific culture medium is used in the process that the myeloid cells are differentiated into macrophages, finally induced and differentiated macrophages are mainly M2 type (accounting for nearly 70%), and tissue regeneration markers are highly expressed on the surfaces of the macrophages. The cartilage regeneration can be promoted; the suspension cells can be collected at a fixed time in a myeloid cell maturation process. According to the invention, the problems of low differentiation efficiency, low purity, long period, immature cell function, dependence on animal-derived components, difficulty in large-scale production and the like in the process of differentiating the pluripotent stem cells into the macrophages are solved.

The purpose of the present invention is to provide tendon / ligament-like artificial tissues with sufficient strength. The present invention provides tendon / ligament-like artificial tissues having sufficient strength, by embedding mesodermal stem cells, mesenchymal stem cells, or such in a gel having strength to withstand tension loading, and culturing the cells while applying tension loading to the gel and while adding cells and gel. Human iPS cells, tissue stem cells, or such can be used for mesodermal and mesenchymal stem cells, and such.

The present application belongs to the technical field of cattle breeding, and provides a method for promoting the development of bovine follicles by injecting long-acting recombinant follicle-stimulating hormone into the urogenital mesoderm. The method comprises injecting an effective dose of long-acting recombinant follicle-stimulating hormone into the urogenital mesoderm of a cow. By injecting long-acting recombinant follicle-stimulating hormone (LArF) into the urogenital mesoderm, slow, continuous and direct action can be achieved, while off-target effects and pain are minimized. In addition, by using the minimum injection volume, the risk associated with the injection operation can be further reduced. In addition, the urogenital mesoderm injection scheme has the advantages of simpler operation, higher safety, less stress, more efficient process and lower cost under the premise of ensuring the follicle size and oocyte number required for oocyte collection.

The application discloses human pluripotent stem cell-derived sclerotome osteogenic / chondrogenic mesenchymal precursor cells, a preparation method and application thereof, and belongs to the technical field of biological medicines, and solves the problems of poor repeatability, low differentiation efficiency and long differentiation time in the prior art. The preparation method comprises the following steps: step 1, original strip differentiation; step 2, precursor segment mesoderm differentiation; and step 3, subculture and sclerotome mesenchymal precursor cell differentiation. The application has the advantages of scientific design, ingenious concept, simple, specific and easy-to-implement starting cell density control. The differentiation process of the application is simple and short. The cells differentiated by the application express related molecular markers of sclerotome osteogenic / chondrogenic mesenchymal precursor cells and have in-vitro chondrogenic and osteogenic differentiation capabilities. The method has high differentiation efficiency and high repeatability. The differentiation efficiency is stably controlled at 99.2% 0±0.7%.