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

The application provides a high-activity iPSC exosome and a preparation method and application thereof, relates to the technical field of biological medicine, and the preparation method comprises the following steps: obtaining induced pluripotent stem cells; performing flow cytometry sorting to obtain an iPSC subpopulation; performing suspension expansion culture on the iPSC subpopulation to collect a cell culture solution; sequentially performing clarification filtration, ultrafiltration concentration and CD9 antibody affinity chromatography purification, and then eluting and collecting an eluent containing exosomes. The induced pluripotent stem cells overcome donor dependence and batch difference. The specific screening of a cell subpopulation that is positive for CD105, CD73 and CD90 and maintains pluripotency eliminates heterogeneity and guarantees the high activity and uniform efficacy of the exosomes. The use of microcarriers for serum-free suspension culture breaks through the yield bottleneck and realizes large-scale production. Finally, the high-purity and high-activity exosomes are efficiently extracted through ultrafiltration and CD9 antibody affinity chromatography purification.

The application belongs to the technical field of biological medicine, and provides a CXCR4 high expression type iPSC-NK cell with enhanced bone marrow and tumor tissue homing ability, and a preparation method and application thereof. The cell takes pluripotent stem cells as starting cells, overexpresses a membrane-bound IL-15 and IL-15RA fusion protein gene and a CXCR4 receptor gene in the pluripotent stem cells; pluripotent stem cells stably expressing the target gene are obtained, and then iPSC-NK cells are obtained through induction differentiation. The membrane-bound IL-15 and IL-15RA fusion protein gene and the CXCR4 receptor gene are targetedly integrated into a safe harbor site of the induced pluripotent stem cell through a gene editing technology, so as to construct an iPSC cell strain stably expressing key proteins; the function-enhanced iNK cell is obtained through induction differentiation, and exhibits excellent bone marrow and various solid tumor tissue homing ability and persistent immune killing activity.

The present invention discloses, in one embodiment, a method of using human induced pluripotent stem cells to generate three-dimensional human organ tissue for therapeutic drug toxicity and discovery⋅. In one embodiment, a high throughput microtiter plate is loaded with both wild type and Rett disease 3D spheroids and exposed to a drug library, and activity is measured and analyzed for disease rescue to wild type cell behavior.

The present disclosure relates to a method for generating induced pluripotent aprotheria stem cells by chemically reprogramming primary aprotheria stem cells. In certain embodiments, the method also includes the step of selecting induced pluripotent elephant stem cells. Additionally, the cells are transfected with at least (i) C4, (ii) C5, (iii) C6, and (iv) SV40LT.

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 present invention relates to the use of corneal cell-derived mitochondria for the prevention or treatment of corneal damage or corneal endothelial cell diseases. It has been confirmed that mitochondria isolated from corneal endothelial cells differentiated from induced pluripotent stem cells (iPSCs) of the present invention alleviate the inflammatory response of corneal endothelial cells in an inflammatory environment, restore the integrity and function of corneal endothelial cells, and, when delivered into heterologous primary corneal endothelial cells, suppress inflammatory responses, particularly inflammation caused by physical damage, and promote the regeneration of corneal endothelial cells. Therefore, mitochondria possessing such effects can overcome the tumorigenic limitations of conventional stem cell therapies, and due to their ease of production and minimal regulatory requirements, they offer excellent cost-effectiveness and broad applicability.

The application discloses a kind of biological hybrid nanoscale enzyme robot pills, with multi-mode regulation intestinal redox homeostasis and dysbiosis in inflammatory bowel disease, steps are as follows: step one, using zinc nitrate, acetylacetone iron and methanol and other drugs synthesis metal organic framework precursor, then calcination obtains iron single atom nanoscale enzyme;Step two, cultivate enrichment long bifidobacterium and human induced pluripotent stem cells, and extract the exosome or outer vesicle membrane of both respectively;Step three, nanoscale enzyme and probiotic vesicle membrane and stem cell exosome are co-extruded, and biological membrane-nanoscale enzyme composite robot is prepared;Step four, the prepared biological membrane-nanoscale enzyme composite particle is encapsulated in magnetic response pill, and engineering biological hybrid nanoscale enzyme robot pill is formed.Engineering biological hybrid nanoscale enzyme robot pill can be targeted to stay in intestinal inflammatory site under the control of rotating magnetic field after entering the intestinal tract of organism, then promote nanoscale enzyme release and enhance enzyme activity under the action of pulse magnetic field, so as to improve the treatment effect.Engineering biological hybrid nanoscale enzyme robot pill has biocompatibility and treatment safety, and has wide application prospect in the field of treating digestive tract inflammation.

This invention provides a method for promoting the differentiation of induced pluripotent stem cells (iPSCs) into three germ layers and its application in differentiation potential detection, relating to the field of biotechnology. The method includes seeding iPSCs into a culture medium containing 5 μM~20 μM E-cadherin activator 1, shaking culture to form embryoids, adding differentiation medium for differentiation culture, obtaining embryoids with activated three germ layer differentiation potential; seeding the embryoids with activated three germ layer differentiation potential into a container coated with a cell adhesion matrix, adding differentiation medium for further differentiation culture, inducing differentiation into three germ layer cells. This invention solves the technical problems in existing technologies where induction of iPSC differentiation into three germ layers via the embryoid pathway results in poor embryoid formation, poor embryoid state, poor long-term maintenance, and poor adhesion after embryoid transfer, leading to poor endoderm differentiation and migration abilities.

The application discloses a kind of blood vessel microfluidic device and method suitable for glycolipid drug test, the device constructs three-dimensional biomimetic microvessel chip using induced pluripotent stem cells, and integrates multiple sensors;Through integrated environmental control module, programmed blood flow loading and drug exposure are realized.Intelligent analysis system includes multimodal intelligent observation module, based on microscope imaging and artificial intelligence speed measurement technology, three-dimensional flow field is inverted;Microcirculation digital twin module reconstructs blood vessels based on medical imaging and constructs calibratable hemodynamic model, for simulation and prediction;And wet-dry interaction module, through two-way interface, realizes "experiment-simulation" closed loop, injects entity data into digital twin to calibrate, and converts the prediction instruction of digital twin into chip control command to drive verification experiment.The system realizes complete technical chain from biomimetic model construction, intelligent observation, digital twin simulation to active experiment optimization.

To provide a method and composition for generating induced non-pluripotent cells differentiated from single-cell induced iPSC (induced pluripotent stem cell) clone lines. [Solution] A method and composition are provided for obtaining functionally enhanced induced effector cells obtained from targeted differentiation of genome-manipulated iPSCs. The iPSC-induced cells provided herein have stable functional genome editing that results in improved or enhanced therapeutic effects. Therapeutic compositions and their use are also provided, comprising functionally enhanced induced effector cells alone or in combination therapy with antibodies or checkpoint inhibitors.

Disclosed herein are methods for obtaining endometrial stromal fibroblast cells from pluripotent stem cells, such as induced pluripotent stem cells. The present disclosure also provides methods of obtaining a three-dimensional, multilayered endometrial tissue composition. Methods of using the cells and tissue compositions in drug screening and therapeutic applications are also provided.

A myogenin-expressing fibroblast-like cell (MEFLC) line and a construction method and a use thereof are provided. The MEFLC line was deposited in the Guangdong Microbial Culture Collection Center (GDMCC) on Apr. 23, 2022 with an accession number of GDMCC NO: 62409. The construction method includes: 1) constructing a pCW-MYOG-T2A-Puro lentivirus; 2) infecting a human induced pluripotent stem cell (iPSC) line with the lentivirus, inducing the expression of an MYOG gene, and screening out a positive monoclonal cell line; 3) subjecting the positive monoclonal cell line to expanded cultivation, during which the expression of the MYOG gene is continuously induced; and 4) changing cultivation conditions, continuously inducing the expression of the MYOG gene, and screening out positive monoclonal cell lines to ensure the purity of MYOG-positive cells until a cell morphology changes significantly into fibroblastoid cells to obtain the MEFLC line.

Provided here in are methods of producing induced pluripotent stem cells (iPSCs) and isolated population of produced induced pluripotent stem cells (IPSCs). Also provided herein are methods of treating a subject in need thereof using the produced iPSCs or pharmaceutical compositions comprising the produced iPSCs.

本发明公开了一种CD34+造血干细胞化学重编程为诱导多能干细胞的方法。所述方法包括：将CD34+造血干细胞分散为单细胞，接种于包被细胞外基质的培养表面，在无血清培养基中依次进行三阶段诱导培养：第一阶段培养基包含丙戊酸、CHIR99021、RepSox、EPZ6438、PY60、OAC1和OAC2；第二阶段培养基包含丙戊酸、CHIR99021、RepSox、反苯环丙胺、TTNPB、Forskolin、OAC1和OAC2；第三阶段培养基包含CHIR99021、Forskolin、Y‑27632、SP600125、OAC1和OAC2。最后挑取形成的iPSC克隆进行扩增培养，获得诱导多能干细胞。本发明针对CD34+造血干细胞优化了小分子组合和诱导时序，全程无病毒、无转录因子、无血清，重编程效率可达3%以上，细胞凋亡率低，可获得具有完整多能性和正常核型的iPSC，适用于临床级细胞制备和再生医学应用。

The application provides a culture method and culture reagent of a heart organoid, and the scheme of the application uses a culture medium with clear components to perform four-stage induction differentiation culture, so that the heart organoid can be constructed from dispersed human induced pluripotent stem cells, the construction process of the heart organoid is simplified, the construction period of the heart organoid is shortened, and the use of the culture medium system with clear components greatly improves the success rate of heart organoid induction, and can increase the success rate of the traditional heart organoid construction method to 80% to 100%.

This method discloses a technique for inducing the directed differentiation of human amniotic mesenchymal stem cells into liver organoids, comprising the following steps: S1, expanding and culturing third-generation human amniotic mesenchymal stem cells; S2, discarding the culture supernatant from step S1 and adding a first induction medium; S3, discarding the culture supernatant from step S2 and adding a second induction medium; S4, discarding the culture supernatant from step S3, adding trypsin to digest the cells, resuspending the cells in a third induction medium, centrifuging, and culturing; S5, after the completion of step S4, discarding the culture supernatant and adding a fourth induction medium, ultimately obtaining functionally mature liver organoids. This induction method avoids the tumorigenicity problems associated with other methods, such as induced pluripotent stem cell induction, making it more suitable for industrialization and clinical application. Furthermore, it uses simple components and has lower technical requirements, thus greatly increasing the feasibility of the experiment and facilitating industrialization.

This invention relates to the field of biomedical materials technology, providing a heparinized dual-growth factor recellularized adipose tissue decellularized scaffold material, its preparation method, and its applications. The preparation method includes: decellularizing adipose tissue to obtain adipose tissue decellularized scaffold; heparinizing the scaffold to obtain a heparinized adipose tissue decellularized scaffold; loading hepatocyte growth factor and vascular endothelial cell growth factor onto the scaffold to obtain a heparinized dual-growth factor adipose tissue decellularized scaffold; seeding induced pluripotent stem cell-derived hepatocytes onto the scaffold, and culturing to obtain a heparinized dual-growth factor recellularized adipose tissue decellularized scaffold (RAT). This invention uses rat inguinal adipose tissue to prepare RAT material, which is low-cost, readily available, has a simple preparation process, and is easy to scale up. The RAT material of this invention can achieve liver tissue damage repair and hepatocyte regeneration, providing a new treatment strategy for acute liver failure.

Improved compositions and methods for producing iPSCs are needed. [Solution] This specification provides a recombinant circular RNA comprising at least one protein-coding nucleic acid sequence, wherein the protein-coding nucleic acid sequence encodes a reprogramming factor (e.g., a transcription factor), and the reprogramming factor is Oct3 / 4, Klf4, Sox2, Nanog, Lin28, c-Myc, or L-Myc, or a fragment or variant thereof. This specification also provides a method for producing induced pluripotent stem cells (iPSCs), comprising contacting somatic cells with at least one of the recombinant circular RNAs described herein and maintaining the cells under conditions that will result in reprogrammed iPSCs.

This invention discloses the application of induced pluripotent stem cell (EPC) derived from induced pluripotent stem cell differentiation in the preparation of stroke therapeutic agents. Through intravenous injection of EPCs into an animal model of stroke, this invention demonstrates that this method can treat stroke by inhibiting atherosclerosis, reducing cerebral infarction levels, promoting angiogenesis in lesioned brain tissue, improving inflammatory responses in lesioned brain tissue, and repairing nerve damage and brain injury. The above research results of this invention provide a new treatment method for stroke in clinical practice.

The present invention relates to a method for directly differentiating Visceral sensory neurons from human-derived induced pluripotent stem cells or a composition for inducing differentiation of Visceral sensory neurons from induced pluripotent stem cells. The present invention relates to a novel protocol that can differentiate induced-Visceral sensory neurons from induced pluripotent stem cells, whereby human-derived Visceral sensory neurons can be reproduced in an in vitro environment. The induced-Visceral sensory neurons differentiated by the method of the present invention can be used in various fields such as drug screening utilizing the gut-neuron- brain axis, and disease models, and can be used to elucidate the pathogenesis of various sporadic neurodegenerative diseases with unclear etiology.

Provided is a cultivation method for enhancing the differentiation efficiency and functionality of NK cells, based on the finding that the differentiation efficiency of NK cells is increased by selecting and differentiating cells exhibiting specific characteristics, wherein the efficiency of NK cell differentiation from induced pluripotent stem cells was enhanced by altering conditions at specific stages of the process of differentiating induced pluripotent cells into NK cells, conditions for the composition of stage-specific media used, and the treatment methods for the composition, and in the light of acquiring functional NK cells as well as such quantitative increase, the method can be advantageously used for qualitative improvement.

This application discloses a serum-free cryopreservation solution for cells and its applications. The serum-free cryopreservation solution comprises: 1-2% recombinant human albumin, 5-10 μM (-)-Blebbistatin, 50-100 μM antioxidant, 5-10% permeable cryoprotectant, and a basic culture medium free of phenol red indicator. This serum-free cryopreservation solution is suitable for the cryopreservation of various cell types, particularly muscle cells, especially human induced pluripotent stem cell (iPSC) differentiated cardiomyocytes. It can significantly improve the survival rate, structural integrity, and effective preservation of the physiological functions of cardiomyocytes after resuscitation. It is applicable to basic scientific research, disease modeling, drug screening, and clinical treatment related to cardiomyocytes.

The present application relates to the technical field of medicine, discloses a kind of spinal cord organoid preparation based on trichostatin A pretreatment, including human induced pluripotent stem cell differentiation obtained human spinal cord organoid, trichostatin A and GelMA hydrogel;Wherein, the trichostatin A is used to carry out pretreatment to the human spinal cord organoid before transplantation, to activate NRF2 / HO-1 signal pathway;And its application in preparing drug for treating spinal cord injury.The application combines TSA pretreatment and hSCOs transplantation for the first time, realizes the double advantages of "drug pretreatment enhances cell therapy", clearly that TSA plays the role of antioxidation and anti-apoptosis through NRF2 / HO-1 signal pathway, significantly enhances the survival ability of hSCOs in injury microenvironment, and has good clinical transformation prospect.

This invention provides a method for producing induced pluripotent stem cells. [Solution] The present disclosure includes the step of producing induced pluripotent stem cells by expressing exogenous nucleic acids encoding proteins OCT3 / 4, SOX2, KLF4, LIN28, and L-MYC, as well as p53-shRNA, in umbilical cord amniotic stem cells under conditions suitable for reprogramming stem cells. The present disclosure also includes the induced pluripotent stem cell population obtainable by this method, the induced pluripotent stem cell population obtained by this method, a pharmaceutical composition containing the induced pluripotent stem cells of the present invention, a method for differentiating the induced pluripotent stem cells of the present invention, a pharmaceutical composition containing the differentiated induced pluripotent stem cells obtained by this method, and a method for treating congenital or acquired degenerative disorders in a subject, comprising the step of administering target cells differentiated from pluripotent stem cells to the subject.

This invention discloses a method for regulating the directed differentiation of induced pluripotent stem cells based on the theory of visceral syndrome differentiation, relating to the field of stem cell and regenerative medicine. The method includes the following steps: S1: Constructing a dual-modal database integrating TCM visceral syndrome differentiation data and pluripotent stem cell differentiation omics data; S2: Constructing a cross-modal sequence dataset based on the dual-modal database, training the cross-modal sequence dataset using a multilayer Transformer-XL model, analyzing the long-distance dependencies among visceral pathogenesis, pluripotent stem cell differentiation pathways, and regulatory factors, and obtaining their correlation network; S3: Analyzing the repair needs of target viscera, and automatically selecting the optimal combination of differentiation-inducing factors corresponding to TCM syndromes based on their correlation network; S4: Achieving the directed differentiation of pluripotent stem cells into functional cells of target viscera based on the optimal combination of differentiation-inducing factors. This invention improves the efficiency of stem cell directed differentiation and the functional adaptability of stem cells, and has clinical application value in medicine.

The present invention provides a method for producing a γδT cell from an induced pluripotent stem cell, wherein the induced pluripotent stem cell is derived from a cell other than an αβT cell.

This application discloses a tissue regeneration and repair device and method based on induced pluripotent stem cells, including a cell delivery component and an integrated sorting and processing module. The integrated sorting and processing module is a sealed cylindrical cavity with standard Luer conical connectors at both ends. The proximal Luer connector of the cavity is sealed and connected to the cell delivery component, and the distal Luer connector is sealed and connected to a minimally invasive delivery needle. This application achieves an integrated intraoperative synchronous processing architecture, integrating residual cell removal, targeted inactivation, real-time quality control, and in-situ delivery into a disposable sealed tubing module. It eliminates the need for large laboratory equipment, completing the entire process within 10 minutes, completely resolving the industry pain point of the disconnect between sorting and clinical delivery in existing systems. It is compatible with routine surgical procedures and has an extremely low operational threshold.

The present invention provides a method for producing a γδT cell from an induced pluripotent stem cell, wherein the induced pluripotent stem cell is derived from a cell other than an αβT cell.

High efficiency methods for producing retinal pigment epithelial cells from induced pluripotent stem cells (iPSCs) are disclosed herein. The iPSCs are produced from somatic cells, including retinal pigment epithelial (RPE) cells, such as fetal RPE stem cells. In some embodiments, the iPSC include a tyrosinase promoter operably linked to a marker. Methods are disclosed for using the RPE cells, such as for treatment. Methods for screening for agents that affect RPE differentiation are also disclosed.

This invention provides a novel reprogramming strategy for constructing induced pluripotent stem cells (hiPSCs), along with its methods and applications, belonging to the field of stem cell and regenerative medicine. The method involves inducing induced pluripotent stem cells (hiPSCs) by inducing somatic cells through three stages of cell culture (Stage I, Stage II, and Stage III). This method develops an improved reprogramming strategy using human cells as raw materials. Compared to traditional methods, the new strategy incorporates an early pluripotency culture medium for 6-10 days during the reprogramming process to obtain hiPSCs. Compared to traditional methods, the hiPSCs generated by the new strategy exhibit significantly improved differentiation efficiency into mesoderm and endoderm lineages and can efficiently differentiate in vitro into functionally stable pancreatic β cells. The induced pluripotent stem cells obtained using this method provide high-quality seed cell resources for disease modeling and drug screening of organs derived from mesoderm / endoderm, showing broad application prospects.