36 results about "Differentiation induction" patented technology

The main objective of this invention is to provide a novel manufacturing method capable of efficiently and directly converting or inducing somatic cells into insulin-producing cells. As an example, a method for manufacturing insulin-producing cells is provided, which involves directly differentiating and inducing insulin production from somatic cells. This method includes a step of culturing somatic cells in a serum-free differentiation-induction medium; or a step of culturing somatic cells in a differentiation-induction medium containing 5 μg / mL or more of insulin. According to this invention, insulin-producing cells with high insulin secretion capacity can be efficiently and directly manufactured from somatic cells. The insulin-producing cells obtained by this invention are useful in regenerative medicine and the like.

The objective is to provide a novel method for improving the efficiency of osteogenic differentiation induction based on immobilized BMP-2. [Solution] A method for stimulating cells with ligand molecules, comprising a stimulation step of applying physical stimulation to the cells and / or ligand molecules, wherein the ligand molecule is bound to or interacts with a receptor protein expressed on the cells and is immobilized on a culture substrate and / or matrix molecule used for culturing the cells, the receptor protein mediates the transmission of extracellular signals to the cells based on its binding to the ligand molecule, and the extracellular signals are enhanced by the physical stimulation.

Provided are a production method of retinal pigment epithelial (RPE) cells that improves differentiation induction efficiency of pluripotent stem cells into RPE cells, and can provide highly pure RPE cells by a simple and easy operation in a short period, a culture method of RPE cells that can stably grow and culture a cell, a toxicity / efficacy evaluation method using RPE cells useful for transplantation therapy, and a therapeutic drug for a retinal disease. The invention relates to a production method of RPE cells, comprising adhesion culture of human pluripotent stem cells using a culture substrate coated with a laminin-E8 fragment, a culture method of RPE cells, comprising adhesion culture of RPE cells using a culture substrate coated with a laminin-E8 fragment, a toxicity or efficacy evaluation method using RPE cells obtained by producing or culturing by the method, and a therapeutic drug for a retinal disease, containing the RPE cells.

This invention discloses a method for virus-free tissue culture using terminal buds of *Prunus armeniaca* rootstock, belonging to the field of plant tissue culture technology. The method includes: selecting terminal buds of *Prunus armeniaca* rootstock for pretreatment, sterilizing them with ethanol and mercuric chloride, and then immersing them in a virus-free pretreatment solution containing ribavirin and triazole nucleoside for virus removal; inoculating the virus-free terminal buds into WPM-based differentiation induction medium to obtain clustered buds; culturing the clustered buds in 1 / 2 MS-based proliferation subculture medium to obtain proliferating buds; inoculating the proliferating buds into 1 / 3 MS-based rooting medium to induce rooting; and finally, after hardening off and transplanting into a vermiculite-garden soil-humus substrate for acclimatization. This invention optimizes the virus removal and tissue culture system based on the characteristics of *Prunus armeniaca* rootstock, achieving a virus removal rate ≥95%, a terminal bud differentiation survival rate ≥85%, a rooting rate ≥90%, and a transplant survival rate ≥88%. This enables rapid and large-scale propagation of virus-free *Prunus armeniaca* rootstock seedlings, solving the problem of low propagation efficiency in traditional methods and providing high-quality rootstock support for the *Prunus armeniaca* industry.

The present invention addresses the problem of providing a novel method for increasing bone differentiation induction efficiency based on immobilized BMP-2.　Provided is a method for stimulating a cell with a ligand molecule, the method including a stimulation step for applying a physical stimulus to the cell and / or the ligand molecule. The ligand molecule binds to or interacts with a receptor protein expressed on the cell, and is immobilized on a culture substrate and / or a matrix molecule used for culturing the cell. The receptor protein mediates the transmission of an extracellular signal to the cell on the basis of binding to the ligand molecule, and the extracellular signal is enhanced by the physical stimulus.

The invention relates to a fixing device for loading low-intensity pulsed ultrasound by taking water as a medium and application of the fixing device in promoting osteogenic differentiation of mesenchymal stem cells. According to the device, a double-base type probe fixing structure consisting of a main body fixing base and an accessory fixing base is arranged in a main body structure; and a culture vessel fixing structure on the inner wall of the main body structure is matched, so that the problems that a probe is easy to shift, the heights of culture vessels are inconsistent and the like in the prior art are effectively solved, and a standardized and high-repeatability loading environment is provided for an LIPUS in-vitro experiment. The device is further based on the device, water is used as an ultrasonic conduction medium to be applied to osteogenic differentiation induction of bone marrow mesenchymal stem cells (BMSCs), the efficient and stable osteogenic stimulation effect can be achieved, and the core defects that in the prior art, a probe makes direct contact with a culture vessel, consequently, cells fall off and are curled, morphological damage is caused, and sound field distribution is uneven are overcome; and meanwhile, the stability and repeatability of an experimental result are remarkably improved.

The application provides a preparation method and application of tremella polysaccharide intelligent hydrogel for inhibiting ferroptosis, and relates to the technical field of biomaterial preparation, wherein the tremella polysaccharide intelligent hydrogel for inhibiting ferroptosis is prepared from methacrylated tremella polysaccharide TrepMA, oxidized tremella polysaccharide O-Trep and ketone thioalcohol diamine TK-NH2. The tremella polysaccharide intelligent hydrogel prepared by the method has a three-dimensional network structure, good mechanical properties and can load growth factors or active molecules to accelerate bone regeneration. In addition, the tremella polysaccharide intelligent hydrogel can inhibit ferroptosis and promote bone defect healing with osteoporosis. The tremella polysaccharide intelligent hydrogel prepared by the method has multiple biological activities, can not only provide certain support for bone regeneration, but also can inhibit ferroptosis, improve local osteoporosis, has antioxidant / anti-inflammatory and osteogenic differentiation induction effects through excellent biocompatibility, biological activity and drug controlled release effect, and can effectively promote bone tissue regeneration.

The present application relates to a kind of porous scaffold for repairing bone defect loaded with black phosphorus active oxygen elimination and slow-release drug and its preparation method;The scaffold includes the amino cation polymer grafted with the phenylboronic acid (PBA) and cyclodextrin (CD) that can eliminate active oxygen, black phosphorus nanosheet (BP) and scaffold material poly (lactic acid-glycolic acid) (PLGA).The porous scaffold loads a large number of black phosphorus nanosheet that can be hydrolyzed into bone repair nutrients, and the porous structure provides sufficient space for cell migration and growth, realizes the good repair of bone.Active oxygen elimination ability, can eliminate active oxygen in diabetic microenvironment.In addition, cyclodextrin can load a variety of drugs, such as osteogenic differentiation induction drug, and realize the slow release of drug.The porous scaffold has good biocompatibility, and the degradation product is harmless to human body, can be used for the tissue repair of bone defect, can be widely applied in the field of biological medicine and tissue engineering, and has been proved to have good repair ability in the diabetic rat skull defect model.

A method of producing macrophages includes a culture step of culturing hematopoietic progenitor cells in the presence of a TREM2 signal activator.

Provided is a method for inducing muscle-derived differentiation of porcine pluripotent stem cells, which does not involve transgenosis and is free of serum addition in the whole course, and also provided is a method for preparing cell cultured meat (CM) based on the muscle-derived differentiation induction method.

The invention discloses an osteogenic differentiation induction medium based on pilose antler polypeptide and an osteogenic differentiation induction method, and belongs to the technical field of biology. The osteogenic differentiation induction culture medium based on the cornu cervi pantotrichum polypeptide contains a cornu cervi pantotrichum polypeptide solution, dexamethasone, L-ascorbic acid, beta-sodium glycerophosphate and a complete culture medium, the cornu cervi pantotrichum polypeptide provided by the invention can greatly improve the osteogenic differentiation effect of mesenchymal stem cells as an additive component of an osteogenic induction culture medium, and is superior to a commercial osteogenic induction culture medium in a certain scene; according to the osteogenic induction differentiation method disclosed by the invention, the defects in the prior art are improved, and the osteogenic differentiation effect of the mesenchymal stem cells can be improved by adopting the pilose antler polypeptide osteogenic differentiation induction culture medium.

The present application provides a novel medium composition for use in inducing differentiation of somatic cells into brown adipocytes. As the present application, for example, there can be mentioned a medium composition for use in inducing differentiation of somatic cells into brown adipocytes, which is characterized by comprising seven components of a thyroid hormone receptor agonist, a glucocorticoid receptor agonist, a phosphodiesterase inhibitor, insulin, an ascorbic acid derivative, albumin, and an antibiotic. According to the present application, differentiation induction of somatic cells into brown adipocytes can be effectively performed. In addition, according to the present application, brown adipocytes can be effectively maintained.

The invention relates to the technical field of modern biomedicine, in particular to a method for promoting differentiation of human umbilical cord mesenchymal stem cells based on an EVs composite material, which comprises the following steps: 1, preparing a double-layer composite scaffold; 2, preparation and characterization of EVs: primary cartilage cell culture, enzymatic digestion of cartilage tissue, digestion termination and cell collection, EVs extraction, and EVs characterization identification; 3, loading the EVs; 4, separation, identification and differentiation induction of the hUC-MSCs; and 5, differentiation effect detection. The method has the characteristics that the differentiation efficiency and stability are good, the EVs delivery efficiency is remarkably improved, the biocompatibility and safety are good, the operation is simple and convenient, the cost is controllable, and the application scene is flexible and wide.

The application discloses a preparation method of a multi-phase "core-shell" biomimetic tissue engineering support, and constructs a multi-phase "core-shell" in-situ biomimetic support to promote the reconstruction of a rotator cuff tendon-bone interface; the support is made of PCL / PEO biological materials and mixed with a plurality of bioactive factor microspheres, the support fiber is a "core-shell" structure, the "shell" contains SDF-1 chitosan slow-release microspheres, the "inner core" is a bone tissue layer, a fibrocartilage layer and a tendon layer, the layers are continuously printed, the spatial distribution of various factors is fine and controllable, the tendon-bone interface is reconstructed in a biomimetic manner, the sequential controllable release of SDF-1 in the outer layer and various differentiation induction factors in the inner core is realized by using the "core-shell" structure fiber, the endogenous MSCs recruitment and the ordered directional differentiation of each layer are promoted, and the application develops an electrohydrodynamic coaxial 3D printing technology on the basis of the electrohydrodynamic printing technology, and constructs a multi-phase "core-shell" in-situ biomimetic support to promote the reconstruction of the rotator cuff tendon-bone interface.

Provided is a method of generating iPS cells. Specifically, provided is a method of generating iPS cells having a rearranged γδ-TCR gene. Also provided is a cell population including the generated iPS cells. The method includes stimulating collected blood cells with IL-2 and a bisphosphonate, and then introducing cell reprogramming factors through use of a Sendai virus (SeV) vector. According to the method of the present invention, iPS cells having a rearranged γδ-TCR gene can be effectively generated. In particular, the method may be free of a step of treating the blood cells with an antibody before the step of stimulating blood cells with any one kind or a plurality of kinds of interleukins selected from IL-2, IL-15, and IL-23, and a bisphosphonate. In addition, iPS cells generated by the method of the present invention can be differentiated into desired cells by differentiation induction treatment.

The present invention addresses the problem of providing a method for simply and efficiently inducing oocytes and prespermatogonium cells to differentiate from primordial germ cell-like cells or primordial germ cells. This problem is solved by a method for producing oogonium cells or prespermatogonium cells, which comprises a step for culturing primordial germ cell-like cells or primordial germ cells in a culture medium containing a BMP receptor type 2 signaling agonist to obtain oogonium cells or prespermatogonium cells.

Prediction of a differentiation level of a cultured cell is performed non-invasively and simply in an early stage from the end of differentiation induction. A prediction method includes collecting the culture supernatant of the prediction target cultured cell after the start of differentiation induction and before the end of differentiation induction, measuring the content of HICA in the culture supernatant of the collected prediction target cultured cell (step S22), and inputting the content of HICA in the culture supernatant of the prediction target cultured cell measured in the prediction model to predict a differentiation level of the prediction target cultured cell (step S23).

The present application provides a method for producing, from pluripotent stem cells, neural progenitor cells of the midbrain floor plate region or dopaminergic neural progenitor cells, the method comprising (1) inducing differentiation of pluripotent stem cells into neural progenitor cells of the midbrain floor plate region, and optionally (2) inducing differentiation of the neural progenitor cells of the midbrain floor plate region, which are obtained by the differentiation induced in (1), into dopaminergic neural progenitor cells, wherein all or part of the differentiation induced in (1) includes culturing the cells in a medium containing an agonist for LPA receptor.

The present application belongs to the field of biomedical materials, and particularly relates to a hydrogel composition with double functions of stem cell recruitment and osteogenic differentiation promotion, and a preparation method and application thereof. The hydrogel composition comprises a first modified hydrogel and a second modified hydrogel which is spatially adjacent to the first modified hydrogel, wherein the first modified hydrogel contains a hydrogel component and a polypeptide, a protein or an active fragment thereof with a stem cell recruitment function which is connected to the hydrogel component by chemical grafting; and the second modified hydrogel contains a hydrogel component and an osteogenic induction active ingredient which is connected to the hydrogel component by chemical grafting. Through the design of "outer recruitment-inner induction", the hydrogel has double functions of efficient stem cell recruitment and osteogenic differentiation induction, forms a complete repair chain, improves the repair efficiency, and improves the producibility and clinical practicability.

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.

An object of the present invention is to provide a cell culture substrate with laminated cardiomyocyte sheets for evaluating a curative effect by biological transplantation of a normal cardiomyocyte sheet and a method for producing the same. A further object of the present invention is to provide a method for evaluating a curative effect of a normal cardiomyocyte sheet for biological transplantation. The cell culture substrate with laminated cardiomyocyte sheets of the present invention includes two or more layers of cardiomyocyte sheets on a cell culture substrate, wherein the cell culture substrate includes an alternating arrangement portion in which a strip-shaped cell adhesion region and a strip-shaped cell adhesion suppression region are alternately arranged, the cell culture substrate with laminated cardiomyocyte sheets includes the two or more layers of cardiomyocyte sheets on at least the alternating arrangement portion of the cell culture substrate; and the cell culture substrate with laminated cardiomyocyte sheets includes, as the cardiomyocyte sheets, at least one layer of each of a disease cardiomyocyte sheet containing a differentiation-induced cardiomyocyte derived from a disease patient and a normal cardiomyocyte sheet containing a differentiation-induced cardiomyocyte derived from a healthy human.

Provided is a γδ T cell for securing the purity and number of cells sufficient for treatment. Also provided is a method of generating the γδ T cell. More specifically, provided are homogeneous γδ T cells excellent in that the γδ T cells are not affected by exhaustion of the cells. The foregoing is achieved by γδ T cells obtained by subjecting induced pluripotent stem cells (iPS cells) to differentiation induction treatment. Specifically, the foregoing is achieved by γδ T cells generated by subjecting iPS cells having a rearranged γδ TCR gene (γδ TCR-type iPS cells) to differentiation induction treatment. According to the method of generating the γδ T cell of the present invention, there can be provided γδ T cells and a cell population of γδT cells that have an excellent function of having antigen-specific cytotoxic activity in an MHC-unrestricted manner, and that are more homogeneous and have a higher effect than γδ T cells separated from peripheral blood.

The invention discloses a surface-engineered iron-nitrogen-selenium monatomic nano-enzyme loaded with all-trans retinoic acid, which comprises an inner core and all-trans retinoic acid loaded on the surface of the inner core, and the inner core is a selenium-rich organic nano-enzyme inner core. The invention also discloses a corresponding preparation method and application thereof. The ATRA is loaded on the surface of the inner core, the inner core can regulate the active oxygen level in target cells so as to cooperate with all-transretinoic acid to induce acute myelogenous leukemia cell differentiation, the regulating effect of the nano-enzyme on ROS and the differentiation promoting effect of the ATRA generate a synergistic effect, the differentiation induction effect on AML cells is remarkably enhanced, and the differentiation induction effect on the AML cells is remarkably improved. The problem of drug resistance of ATRA single-drug treatment is solved. The nano-enzyme disclosed by the invention not only can effectively inhibit proliferation and infiltration of leukemia cells, but also can promote repair of a bone marrow hematopoietic microenvironment and recovery of a hematopoietic function, and provides a targeted redox differentiation enhancement treatment strategy with clinical transformation potential for acute myelogenous leukemia.

The invention belongs to the field of biotechnology medicine, and particularly relates to a method for improving the induction efficiency of transdifferentiation from astrocytes to neurons, which comprises the following steps: preparing primary astrocytes: separating cortical tissues to prepare a cell suspension, and culturing and purifying to obtain the astrocytes. And transdifferentiation induction: transducing the astrocytes by using lentivirus, changing a differential medium and adding BDNF for culture to promote the astrocytes to be transdifferentiated into mature neurons. Animal model and targeted induction: establishing a cerebral infarction model, and injecting AAV-Neurog2-Pax6 virus into a focus area to realize specific transdifferentiation of astrocytes; the method comprises the following steps: performing in-vivo induction and sample treatment: performing perfusion fixation, dehydration, embedding and freezing slicing after taking materials, retaining the integrity of transdifferentiated cells, and performing overexpression on Neurog2 and Pax6 to realize transdifferentiation of the astrocytes to neurons and improve the induction efficiency of transdifferentiation of the astrocytes to the neurons.

The present invention relates to a method for memory B cell-specific differentiation induction and to uses thereof and, more specifically, to an anti-CD3 antibody or ligand in a biological sample obtained from and individual, a method for memory B cell-specific differentiation induction comprising a step of treating an anti-CD28 antibody or ligand, and a method for detection a memory B cell which is specific to a specific antigen by using same.

The invention relates to a duck induced pluripotent stem cell differentiation method which comprises the following steps: (1) establishment of duck iPS cells: separating and culturing adult cells, transfecting transcription factor plasmids, and screening and amplifying iPS cell clone; (2) differentiating the duck iPS cells into PGC cells: preparing an induced differentiation culture medium, and performing differentiation induction on the iPS cells; and (3) cryopreservation and resuscitation of the PGC cells: preparing a cryoprotectant, carrying out cryopreservation on the PGC cells, and resuscitating the PGC cells. According to the method, differentiation of duck iPS cells and preservation and recovery of PGC cells are successfully achieved, genetic resources of endangered duck species can be effectively preserved, population rejuvenation and ecological restoration of the endangered duck species are promoted, and important technical support is provided for global biodiversity protection.

The invention discloses a preparation method of a human induced pluripotent stem cell induced heart organoid, and belongs to the field of biomedical engineering. The method aims at solving the problems that existing heart organoid is low in induced differentiation efficiency, low in myocardial cell purity, immature in organoid structure and function, complex in construction process, poor in repeatability and the like. The method comprises the following steps: culturing and amplifying human induced pluripotent stem cells, and inducing embryoid bodies for 7.5 days by an induction culture medium containing BMP4, FGF-2, CHIR99021, IWP-2, retinoic acid, Insulin and the like after the embryoid bodies are formed to form heart organs. The invention aims to provide a set of detailed and standardized operation process to guide a user to successfully induce and differentiate to generate heart organs with a pulsation function by using the human induced pluripotent stem cells. The method covers the whole process from human induced pluripotent stem cell preparation, stage differentiation induction to heart organoid collection and identification, and can be applied to the fields of heart disease model construction, drug screening and evaluation, regenerative medicine research and the like.

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.

The invention belongs to the technical field of biology, and particularly relates to a culture medium capable of differentiating and inducing amniotic fluid cells into myoblasts, a method and application. The culture medium contains 5% v / v serum, 0.1 [mu] M of dexamethasone, 50 [mu] M of hydrocortisone, 1 [mu] M of MCHIR99021 and 5-10 ng / mL of IGF-1, and a solvent is a low-sugar DMEM (Dulbecco Modified Eagle Medium). According to the invention, amniotic fluid cells are separated from amniotic fluid, and myoblasts can be obtained by using the culture medium to culture, differentiate and induce. According to the invention, efficient muscle differentiation and RYR1 gene overexpression of amniotic fluid cells are realized for the first time, and the gene has high fidelity on splicing variation after induced differentiation, so that a safe, rapid and low-cost solution is provided for prenatal clear gene diagnosis.

This invention provides a method for evaluating stem cell trilineage differentiation based on single-cell membrane tension measurement, comprising: measuring the membrane tension of multiple single cells in a stem cell sample to be tested, and calculating the mean membrane tension; setting a membrane tension threshold for stem cell trilineage differentiation, comparing the mean membrane tension with the threshold, and evaluating the differentiation process and efficiency of the stem cell sample to be tested. This invention also provides the application of the above evaluation method in the quality control of clinical-grade stem cells, the evaluation of stem cell differentiation induction effects, or the screening of stem cell virulence before transplantation. This invention utilizes optical tweezers technology to construct a single-cell mechanical testing platform, enabling non-destructive quantitative measurement of single-cell membrane tension. Cells can remain intact and viable after testing and can be reused. By comparing the membrane tension data of the sample to be tested with the threshold, a systematic evaluation of stem cell differentiation process and efficiency can be completed, which can be widely applied in the fields of stem cell quality control and regenerative medicine clinical translation.