407 results about "Human Induced Pluripotent Stem Cells" patented technology

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 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.

Materials, methods, and systems are provided for cell redifferentiation and expansion of induced pluripotent stem cell (iPSC)-derived hematopoietic stem cells (iHSCs) without the use of serum or additional cells.

The present invention provides methods of producing in vitro derived neutrophils or macrophages in xenogen- and serum-free conditions from pluripotent stem cells and in vitro derived populations of neutrophils and macrophages. Methods of treatment using in vitro derived neutrophils or macrophages are also contemplated.

The invention provides cell culture conditions for culturing stem cells, including feeder-free conditions for generating and culturing human induced pluripotent stem cells (iPSCs). More particularly, the invention provides a culture platform that allows long-term culture of pluripotent cells in a feeder-free environment; reprogramming of cells in a feeder-free environment; single-cell dissociation of pluripotent cells; cell sorting of pluripotent cells; maintenance of an undifferentiated status; improved efficiency of reprogramming; and generation of a naïve pluripotent cell.

The invention relates to the technical field of preparation of organoids, and discloses a method for regulating and controlling human induced pluripotent stem cells to differentiate into kidney organoids, which comprises the following steps: resuspending a cell cluster formed by human induced pluripotent stem cells by using a basic differentiation culture solution containing a GSK-3beta inhibitor, inoculating the cell cluster into an ultralow adsorption cell culture plate, and starting to culture, marking the time as the 0th hour of differentiation; and after culturing for 70-76 hours, replacing the basic differentiation culture solution containing the GSK-3beta inhibitor with a basic differentiation culture solution containing a Knockout TM serum substitute (KSR), and continuing to carry out differentiation culture. By adopting the method disclosed by the invention, the human induced pluripotent stem cells can be efficiently, quickly, simply, conveniently and stably subjected to directional induced differentiation into kidney organs containing podocytes, renal tubules and other structures, and the time for constructing a kidney in-vitro model can be shortened, so that the research on kidney diseases, kidney development and the like can be greatly assisted; and a powerful platform is provided for kidney toxicity assessment.

The present invention provides isolated human breast milk exosomes and compositions thereof. The invention also provides an in vitro method for producing the exosome, comprising generating lactating cells derived from human induced pluripotent stem cells (hiPSCs), expressing a human milk-like product from the lactating cells, and purifying the exosome from the human milk-like product.

A microfluidic 3D-printed hydrogel based on fish liver decellularized extracellular matrix for liver regeneration and a preparation method. The hydrogel is prepared by means of combining fish liver decellularized extracellular matrix (dECM) and gelatin methacryloyl (GelMA), and loading induced pluripotent stem cell-derived hepatocytes (iPSC-heps) for liver regeneration. The microfluidic 3D-printed hydrogel based on fish liver decellularized extracellular matrix exhibits excellent biocompatibility, retains intact endogenous growth factors, maintains the biological activity of cells, ensures the effective encapsulation of the cells, and facilitates the robust functional expression of the iPSC-heps. After in-vivo transplantation, the survival rate and liver function of mice with acute liver failure are significantly improved, and liver regeneration and repair are promoted.

Injectable compositions and methods of preparation, as well as therapeutic uses, of induced pluripotent stem cell (iPSC)-derived notochordal cell (iNC)-loaded microgels are provided. Microfluidic on-chip platform can be utilized to prepare microgels (or microgel particles / spheres) formed from block copolymers that exhibit reverse thermal gelation, so as to encapsulate iNCs. Also provided are preconditioned iNC-loaded microgels and iNCs in bulk hydrogel. Cell purity, identity, viability, sterility, and the stability of microencapsulated iNCs have been evaluated. Safety and efficacy of the compositions as therapeutic candidates has been tested via intradiscal injection in animal models of intervertebral disc (IVD) degeneration and discogenic low back pain. Biobehavioral testing, MRI, and immunohistochemical analyses were utilized to evaluate the regenerative potential and reproducibility of the compositions as therapeutic candidate. Single cell RNA sequencing of the treated IVDs may also reveal mechanism of action of the compositions.

The present disclosure provides engineered induced pluripotent stem cells (iPSCs) expressing B2M-HLA fusion constructs. The present disclosure also provides iPSC-derived macrophages which overexpress IL-12. The present disclosure further provides engineered immune cells having persistent cytokine expression.

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.

The present invention discloses a method for establishing and applying a caudal fin-induced pluripotent stem cell line of Erythroculter ilishaeformis. By the tissue adherence method, a cell line with high proliferation performance and stemness is isolated from the caudal fin of Erythroculter ilishaeformis. In the early stage, the cells of this cell line are significantly fibroblast-like. Later, the cells become smaller but still maintain a spindle shape, without obvious contact inhibition phenomenon, and can spontaneously form cell clusters similar to induced pluripotent stem cells. Alkaline phosphatase staining is positive and it expresses stemness marker genes such as Oct4, Sox2, Klf4, Nanog, and c-Myc. The present invention uses a single cAMP agonist small molecule compound to reverse the phenomenon of weakened stemness after continuous subculture of this cell line, enabling it to maintain the characteristics of high proliferation performance and high stemness, which is suitable for future research on the induction and differentiation of fish stem cells and provides more choices for the research and application of fish stem cells.

This disclosure relates to cell therapy approaches for treating progranulin (PGRN) deficiencies with human induced pluripotent stem cell (hiPSC)-derived cells. Advantageously, the hiPSC-derived cells (e.g., microglia progenitor cells) described herein can cross-correct PGRN deficiencies in damaged or diseased cells while reducing the amount of endogenous cell ablation that is needed, as demonstrated by experimental results showing restoration of PGRN levels in GRN mutant cells and brain organoids.

The present disclosure provides efficient in vitro methods for producing megakaryocytes and / or platelets from human induced pluripotent stem cells (iPSCs). In various embodiments, the cells and platelets produced in accordance with the present disclosure are functional and / or more closely similar to corresponding lineages isolated from peripheral blood, bone marrow, or other tissue. The present invention provides, in some aspects, isolated cells / platelets and compositions produced by the methods disclosed herein, as well as methods for use in therapy.

This invention relates to the technical field of stem cell induction differentiation and transplantation therapy, specifically to a method for inducing human induced pluripotent stem cells to differentiate into dopaminergic neuronal progenitor cells, comprising the following steps: S1, human induced pluripotent stem cells are induced to differentiate into primitive neuroepithelial cells under the action of SB431542, DMH1, SHH, FGF8b, Purmorphamine, and CHIR99021; S3, intermediate primitive neuroepithelial cells are continuously induced to differentiate using BDNF, GDNF, AA, N2, B27 (without added vitamin A), cAMP, and GlutaMax to obtain dopaminergic neuronal progenitor cells. The dopaminergic neuronal progenitor cells obtained using the method provided by this invention can survive for a long time in the brain microenvironment of Parkinson's rats and have a significant repair effect on rat motor dysfunction.

It is revealed that an organ such as pancreas can be regenerated by utilizing a fact that the deficiency of an organ is complemented by injecting an induced pluripotent stem cell (iPS cell) into a developed blastocyst in a blastocyst complementation method. Thus, the present invention has solved the above-described object. This provides a method for producing a target organ, using an iPS cell, in a living body of a non-human mammal having an abnormality associated with a lack of development of the target organ in a development stage, the target organ produced being derived from a different individual mammal that is an individual different from the non-human mammal.

The invention belongs to the field of cell biology, and particularly relates to a preparation method and application of an extracellular matrix of cell-engineered human collagen. The preparation method comprises the following steps: S1, culturing human induced pluripotent stem cells or a cell culture comprising the human induced pluripotent stem cells, and directly inducing differentiation to obtain induced mesenchymal stem cells or a cell culture comprising the induced mesenchymal stem cells; and S2, carrying out 3D culture on the induced mesenchymal stem cells through a 3D porous scaffold carrier. The extracellular matrix prepared by the preparation method provided by the invention is rich in collagen, so that the mesenchymal stem cells can be better promoted to be differentiated into osteoblasts, and meanwhile, a new thought is provided for the preparation of the collagen.

The present disclosure provides, in various aspects and embodiments, methods for generating hematopoietic lineages for cell therapy, including erythroid progenitor cells, progenitor cell erythroblasts, granulocyte-macrophage progenitor cells (GMP), and megakaryocyte erythroid progenitor cells (MEPs) and erythroid cells. In various embodiments, the invention provides efficient in vitro methods for developing such hematopoietic lineages, including but not limited to progenitor erythroblasts and erythroblasts lineages, from human induced pluripotent stem cells (iPSCs). The cells produced in various embodiments according to the present disclosure are functional and / or more closely similar to corresponding lineages isolated from peripheral blood or bone marrow. The invention also provides isolated cells and cell compositions produced by the methods disclosed herein, as well as methods for cell therapy.

The present invention relates to a method for three-dimensional culture and induced differentiation of neural organoids and use thereof. The method for three-dimensional culture and induced differentiation of neural organoids comprises: adding a signal pathway inhibitor into an induced pluripotent stem cell three-dimensional culture, and culturing cells to obtain the neural organoids. The signal pathway inhibitor comprises a TGF-β signal pathway inhibitor, a WNT signal pathway inhibitor, and a BMP signal pathway inhibitor. Compared with a traditional method, the neural organoids obtained by the method of the present invention feature a large size, small individual difference, and short culture time. The method is simple and rapid, and has high stability and low cost, and only a common incubator is needed. The method has wide application prospects.

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

The invention provides a method for promoting differentiation of induced pluripotent stem cells to trigerm layers and application of the method in differentiation potential detection, and relates to the technical field of biologication.The method comprises the steps that the induced pluripotent stem cells are inoculated into a culture medium containing 5-20 [mu] M of E-cadherin activator 1, shaking culture is conducted to form embryoid bodies, the embryoid bodies are added into a differentiation culture medium for differentiation culture, and the embryoid bodies are obtained. Obtaining an embryoid body for activating three-germ-layer differentiation potential; inoculating the embryoid body which activates the differentiation potential of the three-germ layer into a container coated with a cell adhesion matrix, adding a differentiation culture medium, continuing differentiation culture, and performing induced differentiation to form the three-germ layer cells. The technical problems that in the prior art, when iPSC is induced to be differentiated to three germ layers through an embryoid body path, the embryoid body forming capacity is poor, the embryoid body state is poor, the long-term maintaining capacity is poor, and the endoderm differentiation and climbing-out capacity is poor due to the fact that the embryoid body is poor in wall attaching capacity after plate rotation are solved.

The present invention provides methods and compositions for the generation of microglial progenitor cells and microglial cells from pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem cells. The present invention also provides cells produced using such methods, and both methods of treatment and methods of drug screening that use such cells. Also provided are various tissue culture media, tissue culture media supplements, and kits useful for the generation of human microglial progenitor cells and human microglial cells.

The present invention belongs to the technical field of cell biology, and particularly relates to the application of Hippo-Yap pathway regulators in promoting the directed differentiation of induced pluripotent stem cells into pancreatic progenitor cells. The method of the present invention can efficiently obtain pancreatic progenitor cells derived from non-human primate rhesus monkeys in vitro, providing a material source for further obtaining functional pancreatic islet β cells of non-human primate rhesus monkeys, and more importantly, providing a practical basis for the autologous transplantation treatment of non-human primate diabetic animal models.

There is provided a mutant KLF protein that can induce reprogramming of a somatic cell at a higher efficiency than a KLF protein having a natural amino acid sequence. There is also provided a method for efficiently producing an iPS cell by using the mutant KLF protein. There is provided a mutant KLF protein having an amino acid substitution, or a peptide fragment thereof containing the amino acid substitution.

The present invention provides a production method for iPS cells that uses a cell processing device. The cell processing device has a closed system portion at which a sealed container for material supply and a centrifugation chamber for counterflow centrifugation are connected by a connection conduit. The production method involves performing the following steps in order while keeping the closed system portion closed: a step (s1) for bringing an initialization factor into contact with somatic cells while performing counterflow centrifugation in the centrifugation chamber; and a step (s2) for establishing iPS cells from the somatic cells.

The invention discloses a method for constructing astrocytes serving as a smoke disease model, and belongs to the technical field of crossing of stem cells and neuroscience. The method comprises the following steps: S1) reprogramming CD34 + cells in in-vitro PBMCs (peripheral blood mononuclear cells) of smoke disease patients carrying and not carrying RNF213p.R4810K mutation to obtain induced pluripotent stem cells; and S2) directionally inducing and differentiating the induced pluripotent stem cells into astrocytes through a neural progenitor cell way, wherein the obtained astrocytes are the astrocytes capable of being used as the smoke disease model. The astrocyte model prepared by the invention can be used for researching pathogenesis, nerve-blood vessel interaction process and blood-brain barrier (BBB) related functions of smoke diseases, and can be further applied to molecular typing of diseases and in-vitro function evaluation of candidate drugs.

Novel methods for producing self-renewing erythroblasts (SREs) from induced pluripotent stem cells (iPSCs) or CD34+ cells are provided. Also provided are genetically modified iPSCs or CD34+ cells having a constitutively active stem cell factor (SCF) receptor and a constitutively active Janus kinase 2 (JAK2) that can differentiate into enucleated red blood cells, self-renew, and expand in culture media for an extended period of time. In particular, the disclosed methods completely eliminate the need for SCF and Epo.

The present invention relates to methods of determining a potency state of an embryonic stem cell (ESC) in culture, comprising detecting the expression level of Negative Elongation Factor Complex Member A (Nelfa) in the ESC, and correlating the expression level of Nelfa with that of a reference sample to determine the potency state of the ESC. The present invention also provides methods of selecting totipotent-like ESCs from a population of ESCs in culture using an anti-Nelfa antigen binding protein, methods of inducing totipotency in an ESC in culture, methods of improving the reprogramming efficiency of a somatic cell into an induced pluripotent stem cell and methods of reprogramming a somatic cell into a totipotent stem cell by inducing Nelfa expression. Cells comprising an expression vector encoding Nelfa and kits for use in the methods of the invention are also provided.

Provided herein are methods and compositions for differentiating induced pluripotent stem cells into granulosa-like cells by overexpressing transcription factors such as NR5A1 and a RUNX family protein (e.g., RUNXI and / or RUNX2).