524 results about "Pluripotential stem cell" patented technology

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

The invention discloses application and a preparation method of exosome EGF-FGF-EXO containing double growth factors, the exosome EGF-FGF-EXO is uniform in particle size, high in loading efficiency and good in biological activity, the double growth factors comprise an epidermal growth factor EGF and a fibroblast growth factor FGF, and the epidermal growth factor EGF and the fibroblast growth factor FGF are added into the exosome EGF-FGF. The preparation method of the exosome EGF-FGF-EXO comprises preparation of an induced pluripotent stem cell (iPSC) exosome and loading of EGF-FGF, a recombinant epidermal growth factor (EGF) and a fibroblast growth factor (FGF) are loaded on the exosome from iPS-clone 12c, and the exosome product which has high biological activity and is used for treating nervous system diseases of neuroinflammation is prepared. The invention develops a brand new technical path for treatment of nervous system diseases, and in the fields of epilepsy, neurodegenerative diseases, brain injury and the like, the life quality of patients can be improved and important theoretical basis and technical support can be provided for precise medical treatment of nervous system diseases by precisely regulating and controlling neuroinflammation.

The invention belongs to the technical field of organoid culture, and particularly relates to a co-differentiated vascularized liver organoid and a construction method thereof. Aiming at the problems that the existing liver organ vascularization mostly adopts a co-culture mode, the operation process is complicated, the cell source heterogeneity is relatively high, the randomness is strong, the uniformity is poor and the like, the invention provides the construction method of the co-differentiated vascularized liver organ. The preparation method comprises the following steps: sequentially inducing and culturing directional endoderm cells, posterior preintestinal cells and hepatic endoderm cells, adding an S1 culture medium containing VEGF to induce and culture for 1-3 days, then adding an S2 culture medium containing VEGF, BMP4 and FGF2 to induce and culture for 3-5 days, and then adding an S3 culture medium containing VEGF to induce and culture for 14-20 days to obtain the co-differentiated vascularized liver organ. According to the method disclosed by the invention, the operation process is simplified, the experiment efficiency is improved, the liver organ which is rich in blood vessel network, high in maturity and of a blood vessel-bile duct structure is obtained, and a foundation is laid for large-scale production and clinical application.

The invention provides a bone organ model as well as a construction method and application thereof, and the method comprises the following steps: arranging a biological material with a micropore array in a cell culture plate to obtain a bone organ chip; suspending the initial stem cells in a culture medium, and promoting the cells to form a spherical structure through cell aggregation or by using a non-adhesive culture surface, so as to obtain cell spheres; and culturing the cell spheres in the bone organ chip by adopting the culture medium for constructing the bone organ model to obtain the bone organ model. The skeletal organ is successfully constructed through in-vitro self-organized 3D co-culture initial stem cells including embryonic stem cells, induced pluripotent stem cells, bone marrow mesenchymal stem cells, osteoblasts, bone cells, all levels of osteogenic precursor cells, osteogenic-like cells and MC3T3-E1 cells, and the skeletal organ can become an important tool for researching and treating bone related diseases.

The invention discloses a 3D intestinal organ differentiation method based on human pluripotent stem cells and an induction culture medium and application thereof, and relates to the technical field of stem cells. According to a culture medium formula combination, intestinal organs can be differentiated into various cell types such as epithelial cells, neuroendocrine cells and endothelial cells; the method is a key mark for successful differentiation and functional maturation of intestinal organs. According to the 3D intestinal organ differentiation method disclosed by the invention, histological structures such as intestinal crypts are differentiated from intestinal organs generated by differentiation, and the intestinal organs can creep in a maintenance stage, so that the intestinal organs are changed from structural bionics to functional simulation, and the significance of the 3D intestinal organ differentiation method is far better than that of pure morphological simulation. Through a systematic culture medium formula, a clear operation process and a multi-stage induction strategy, the 3D intestinal organ with structural integrity, cell diversity and functional activity is successfully constructed, and the system provides an efficient, reliable and extensible in-vitro model platform for intestinal biological research and related application.

Embodiments of the invention include compositions and methods related to engineered human mucosal-associated invariant T (eMAIT) cells for off-the-shelf use for clinical therapy for cancer, infectious, and autoimmune diseases. In some embodiments, the eMAIT cells are produced from healthy human donor peripheral blood, cord blood, or G-CSF mobilized peripheral blood. In particular embodiments, the eMAIT cells are produced from a pluripotent stem cell line and therefore can be of unlimited supply. In some embodiments, the eMAIT cells are engineered to express chimeric antigen receptors (CARs), or / and immune regulatory molecules, or / and allorejection resistance molecules. Embodiments of the invention also include compositions of matter comprising polynucleotides encoding mucosal-associated invariant T cell receptor alpha chain polypeptides and / or mucosal-associated invariant T cell receptor beta chain polypeptides.

The present invention relates to pluripotent stem cell comprising an expression construct for expression of a CEBPα protein and an expression construct for expression of an ADIG protein. The invention further provides for methods of producing adipocytes comprising the pluripotent stem cells and for foodstuff comprising the adipocytes or pluripotent stem cells.

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.

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 construction method and application of thymus organoid, and the method comprises the following steps: differentiating pluripotent stem cells into thymus organoid under a suspension culture condition through combined and matched use of small molecule compounds and small molecule proteins, according to the method disclosed by the invention, the thymus organoid of which the structure and the function are closer to those of natural thymus can be efficiently obtained.

The invention provides a pancreatic islet organ capable of efficiently maintaining pancreatic islet beta cell identity and a preparation method and application thereof. Specifically, the invention provides the preparation method for the in-vitro pancreatic islet organ, and the preparation method comprises the following steps: carrying out induced differentiation culture on the ZnT8-knocked-out pluripotent stem cells, so as to obtain the mature pancreatic islet organ. The invention also provides a corresponding transplantation composition and application thereof. The invention discloses that ZnT8 knockout is a key factor for effectively maintaining the identity stability of beta cells in pancreatic islet organs. In addition, the pancreas islet beta cell identity maintenance agent is combined with the ZnT8-knocked-out pancreas islet organ for use, so that the identity stability of the pancreas islet organ beta cell can be obviously maintained. The invention provides a new strategy for transplantation treatment of type 2 diabetes, and has excellent clinical transformation value.

The present invention discloses a method for enhancing cardiomyocyte production from induced pluripotent stem cells (iPSCs) by adjusting the culture medium exchange process during the transition from WNT activation to inhibition for differentiation. Instead of completely replacing the culture medium, this innovative technique involves partially changing the medium, which allows for a controlled exposure to WNT activators concurrent with the introduction of WNT inhibitors. This key modification in the protocol markedly increases the consistency and efficiency of cardiomyocyte generation, reducing variability between batches. By fine-tuning the medium exchange process, this method streamlines the creation of cardiomyocytes for use in regenerative medicine, emphasizing the critical adjustment phase within the culture conditions.

The present invention relates to pluripotent stem cell comprising an expression construct for expression of a Myo1B protein. The invention further provides for methods of producing adipocytes comprising the pluripotent stem cells and for foodstuff comprising the adipocytes or pluripotent stem cells.

The invention belongs to the technical field of medicines, and discloses application of exosomes derived from human pluripotent stem cells in preparation of anti-hepatic inflammation medicines. According to the invention, the exosome derived from the human pluripotent stem cells is applied to the preparation of the anti-hepatic inflammation medicine, especially the preparation of the anti-hepatic fibrosis anti-inflammatory medicine. The human embryonic stem cell-derived exosome can inhibit and regulate polarization of liver macrophages, inhibit inflammatory liver macrophages, up-regulate anti-inflammatory liver macrophages, reduce secretion of inflammatory factors, improve secretion of anti-inflammatory factors and inhibit liver inflammations and development thereof, has an obvious anti-inflammatory treatment effect on inflammation of liver fibrosis, and can be used for preparing medicines for treating liver fibrosis. Therefore, the progress of hepatic fibrosis can be inhibited.

The invention discloses a biconvex crystalline lens organ relying on stiffness-controllable hydrogel external directional induction as well as a construction method and application thereof. A crystalline lens organ model provided by the invention is jointly realized by virtue of stiffness-controllable hydrogel and a composite growth factor; and the pluripotent stem cells are rapidly and directionally induced into crystalline lens organs with high quality in vitro. According to the technical scheme, the method comprises the following steps: (1) directionally inducing and differentiating the pluripotent stem cells into primary ectoderm cell clusters; (2) preparing hydrogel with specific rigidity and carrying out surface cell adhesion treatment; (3) planting the primary ectoderm cells on the cleaned polyacrylamide hydrogel and inducing the primary ectoderm cells into a neural ectoderm cell cluster; and (4) inducing the neural ectoderm cells to directionally differentiate into mature crystalline lens organs. The three-dimensional lens organ-like model obtained by the method has a sphere-like biconvex morphology with a top surface and a bottom surface, is transparent and has structures such as capsule membranes, lens epithelial cells and lens fibrocytes, and the bottom surface is also wrapped by a complete lens capsule membrane. Compared with the prior art, the method realizes the formation of a complete capsule film on the bottom surface and a sphere-like form. The obtained crystalline lens organ can be used for research on crystalline lens embryonic development, research on cataract pathogenesis and research, development and screening of cataract drugs.

The invention relates to a cartilage stem cell for promoting joint repair and application thereof. In particular, the present invention provides a cellular preparation for alleviating or treating osteoarthritis. The cell line comprises Procr-positive pluripotent stem cells. The novel cartilage stem cell provided by the invention can be differentiated in vivo to generate downstream cartilage cells, and cartilage repair in osteoarthritis injury is promoted; it is prompted that selection of Procr positive cartilage stem cells for cell treatment will be a more effective treatment method. And the Procr protein can be used as a new target of the medicine for treating osteoarthritis.

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.

A method for efficiently inducing muscle stem cells to obtain muscle fibers or skeletal muscle. The method comprises: firstly, using a suspension culture and three types of muscle stem cell induction culture mediums to carry out step-by-step induction; then using a culture medium containing bovine serum and a basic fibroblast growth factor BFGF for amplification; then using differentiation culture medium containing specific chemical small molecules for inducing differentiation. The method can be used for rapidly inducing pluripotent stem cells to obtain muscle stem cells and mature muscle fibers or skeletal muscle, and one process can obtain a significant amount of muscle stem cells for treatment and disease model simulation, and thus solving the problem that human muscle stem cells are difficult to obtain and amplify.

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 present invention generally relates to compositions and methods useful for treating a brain injury such as stroke, optic neuropathy, traumatic brain injury, and cerebral palsy. The methods include administering HMCs obtained by in vitro differentiation of pluripotent stem cells and / or extracellular vesicles (EVs) derived from such HMCs (HMC-EVs) into a subject.

The invention provides a method for promoting hematopoietic differentiation of stem cells and application of the method. The method comprises the following steps: carrying out hematopoietic differentiation culture on the stem cells by adopting culture mediums containing histone deacetylase inhibitors in sequence; the culture medium containing the histone deacetylase inhibitor comprises a culture medium I, a culture medium II and a culture medium III, wherein the culture medium I, the culture medium II and the culture medium III are culture mediums containing a histone deacetylase inhibitor; the histone deacetylase inhibitor is selected from sodium butyrate or sodium valproate. The histone deacetylase inhibitor is added into the culture medium for hematopoietic differentiation, so that the efficiency of obtaining the hematopoietic precursor cells through hematopoietic differentiation of the human pluripotent stem cells can be remarkably improved, and hematopoietic differentiation can be effectively promoted.

The present invention relates to a method for inducing the differentiation of corneal epithelial cells from pluripotent stem cells. More specifically, the present invention relates to a method for autonomously differentiating pluripotent stem cells, such as human iPS cells, into ectodermal cell lineage in a serum-free medium without using feeder cells and inducing the differentiation of the resultant ocular surface ectodermal lineage cells into corneal epithelial cells.

Disclosed herein are organoid models of the blood-brain barrier, optionally having a disease associated feature, such as a cerebral cavernous malformation (CCM)-like feature, compositions comprising the same, methods for making the same from pluripotent stem cells, and methods of using the same. These organoids exhibit the complex organization of cells including neurons, endothelial cells, glial cells, and pericytes resembling the natural blood-brain barrier structure. The organoids may also exhibit disease like characteristics, such as having a cerebral cavernous malformation (CCM)-like features. These organoids may be used for studying the functions of the blood-brain barrier and cerebrovascular disorders.

The present invention relates to a method for preparing cardiac organoids derived from human pluripotent stem cells, and an efficacy and toxicity evaluation technique. When the cardiomyocytes and cardiac organoids purified by a CD47 surface marker, of the present invention, are used, mature cardiomyocytes and cardiac organoids can be formed, and the cardiomyocytes and cardiac organoids are characterized in that mature cardiomyocytes having uniform contractile ability and a large surface area are formed at a high rate, and thus the present invention can be effectively used in cardiovascular disease modeling including that in toxicity evaluation and drug screening.

A method for producing mammary gland cells and a method of producing a mammalian milk like product, for example a human milk like product comprising generating lactocytes derived from mammalian induced pluripotent stem cells (miPSC), for example human induced pluripotent stem cells (hiPSC), and expressing the mammalian milk like product, for example the human milk like product from lactocytes.

The present invention provides methods of constructing vascularized retinal organoids by differentiating retinal progenitor cells and endothelial cells. The method is co-cultured by differentiating retinal progenitor cells and endothelial cells, and comprises the following steps: carrying out multiplication culture on pluripotent stem cells until the cell confluence degree is 80-90%, and digesting; carrying out differentiation culture on the digested pluripotent stem cells in a culture medium containing retina induction factors to obtain retina progenitor cells; and co-culturing the retinal progenitor cells differentiated for 12-13 days and endothelial cells to obtain the vascularized retinal organ, wherein the endothelial cells are obtained by any one method of induced differentiation of digested pluripotent stem cells or culture of endothelial cell lines. The vascularized retinal organ with the retinal layered structure is constructed by integrating the endothelial cells and the retinal progenitor cells for the first time, and the limitation that in the prior art, the retinal organ lacks vascularization is successfully overcome.

The invention discloses a stem cell-derived function-enhanced mesenchymal stromal cell as well as a preparation method and application thereof. The preparation method comprises the following steps: inserting an IL-10 gene coding sequence and a CD47 gene coding sequence into a human pluripotent stem cell; a neural crest cell differentiation culture medium containing a Wnt pathway agonist and an ALK5 inhibitor is used for carrying out induced differentiation culture on the CD47 positive human pluripotent stem cells to obtain the neural crest cells. The neural crest cells are subjected to induced differentiation culture by adopting a transition culture medium containing an epidermal growth factor, a fibroblast growth factor 2 and a cell apoptosis inhibitor and a mesenchymal stromal cell differentiation culture medium containing the epidermal growth factor, the fibroblast growth factor 2 and a platelet growth factor in sequence; according to the present invention, with the combination of the specific embodiments, the obtained mesenchymal stromal cell highly expresses CD47 and IL-10, the expression of the CD47 can help the transplanted cell escape from the attack of the immune system and prolong the cell survival time, and the expression of the IL-10 can be used for anti-inflammation.

The present invention discloses a method for culturing vitreoretinal organoids in vitro, comprising the following steps: (1) amplifying and culturing pluripotent stem cells to a cell confluence of 50% to 90%; (2) on day 0, digesting the stem cells with a digestion solution; adding a neurosphere culture medium for culturing; adding matrigel for culturing, and then adding a neurosphere culture medium for culturing to obtain a cell suspension; (3) adding a ROCK pathway inhibitor to the cell suspension, and culturing in the neurosphere culture medium for 3 to 5 days to obtain ectoderm neurospheres; (4) replacing the neurosphere culture medium with a vitreous culture medium I, adding TGFB1 protein, and placing the cell suspension on a shaker for 25 to 28 days; (5) replacing the vitreous culture medium I with a vitreous culture medium II, and culturing the cell suspension for 60 to 300 days to obtain vitreoretinal organoids. The vitreous organoids of the retina cultured in vitro according to the present invention can simulate the vitreous in vivo.