87 results about "Differentiation induction" patented technology

The invention discloses a preparation method of cells with LTo-like functions, which is characterized in that adipose tissue-derived stem cells are treated by LTo differentiation induction signal molecules to obtain cells with LTo-like phenotypes, secretion of cytokines and chemotactic factors of the cells is up-regulated, and the cells have recruitment and support capabilities on lymphocytes in vivo and in vitro. The problems that LTo cells are rare and difficult to obtain are solved, and the adipose-derived stem cells are higher in efficacy, easy to obtain and culture in vitro, capable of being massively amplified, low in cost, efficient and safe.

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 present invention relates to a method for manufacturing a container, its manufacturing apparatus, a cell culture method, a cell culture container, and its manufacturing method and manufacturing apparatus. The present invention provides a container that reduces the risk of contamination and also enables efficient processing such as culturing and differentiation induction of cells in the same container. In addition, a cell culture container capable of ensuring a wide and flat culture surface, etc. is provided. The method for manufacturing a container includes: placing a bag-shaped membrane container on a placement table formed with a recess, introducing a fluid into the interior of the placed bag-shaped membrane container, heating at least one of the placement table and the pressing member, and pressing the bag-shaped membrane container placed on the placement table with the pressing member. On the other hand, the cell culture container includes: a first container wall having air permeability, serving as a bottom surface, and formed of a planar membrane; and a second container wall having a bulging shape that contacts the peripheral portion of the first container wall and protrudes relative to the first container wall; the first container wall is planar at least in a region other than the region in contact with the port.

Provided are: a kit for inducing differentiation, which is for inducing the differentiation of a Muller glial cell into a retinal ganglion cell in vivo in a mammal; and a pharmaceutical composition for treating glaucoma or optic neuropathy. The kit and the pharmaceutical composition each include: a nucleic acid encoding NEUROG2; a combination of a nucleic acid encoding ASCL1, a nucleic acid encoding BRN3B, and a nucleic acid encoding ATOH7; or a combination of a nucleic acid encoding ASCL1, a nucleic acid encoding BRN3B, and a nucleic acid encoding ISL1.

The present disclosure pertains to a method for producing a three-dimensional mammary gland model that comprises a three-dimensional tissue body including mature adipocytes and mammary epithelial cells, the method comprising: a step for gelling a mixture that contains a fragmented extracellular matrix component, an alginate gel precursor, and cells that include adipose-derived stem cells and mammary epithelial cells; and a step for subjecting the gelled mixture to differentiation induction culture to thereby differentiate at least a portion of the adipose-derived stem cells into the mature adipocytes to form the three-dimensional tissue body.

Provided is a method for inducing the differentiation of pluripotent stem cells which are obtained by suspension culture with high quality and / or high efficiency.　The present invention pertains to a method for producing differentiated cells, the method comprising: a step in which thermal stimulation is applied to pluripotent stem cells after suspension culture of the pluripotent stem cells; a step in which the pluripotent stem cells are subsequently seeded in a culture medium that is different from a culture medium used in the suspension culture; and a step in which agregate mass formation and differentiation induction of the seeded pluripotent stem cells are performed.

The invention relates to the technical field of plant tissue culture, in particular to a Shannong crisp pear tissue culture and rapid propagation method which comprises the following preparation steps: S1, selecting top tender shoots of Shannong crisp pears, cleaning and disinfecting the top tender shoots, and inoculating the top tender shoots into a differentiation induction culture medium for culture to obtain cluster buds; s2, transferring the buds into a proliferation culture medium for proliferation culture, then transferring the buds into a rooting culture medium, and inducing rooting. By optimizing the components of each culture medium, the Shannong crisp pear plants are bred, excellent nursery stocks can be quickly obtained, the seedling culture time is effectively shortened, and the seedling culture cost is reduced.

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.

While there have been reports of cultivating islet-like insulin-producing cells by inducing differentiation of ES / iPS cells, the technology for cultivating high-functioning, large-scale insulin-positive islet cells is not yet mature, and there are concerns about rejection and unexpected immune responses. The present invention provides islet-like cells into which the Mycl gene has been introduced, as well as methods for inducing the proliferation of islet-like cells and their differentiation into insulin-producing cells through transient expression of the Mycl gene.

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.

The invention relates to the technical field of plant tissue culture, in particular to a cherry saline-alkaline tolerant rootstock tissue culture and rapid propagation method which comprises the following preparation steps: selecting top tender shoots of cherry saline-alkaline tolerant rootstocks, cleaning and disinfecting the top tender shoots, and inoculating the top tender shoots into a differentiation induction culture medium for culture to obtain cluster buds; and inoculating the cluster buds into a proliferation subculture medium, carrying out proliferation culture to obtain proliferation buds, inoculating the proliferation buds into a rooting medium, and inducing rooting. By optimizing the components of each culture medium and breeding the saline-alkaline tolerant cherry rootstock plants, excellent nursery stocks can be quickly obtained, the seedling culture time is effectively shortened, and the seedling culture cost is reduced.

Provided is a method for inducing differentiation into nerve cells, the method comprising: a) culturing mesenchymal stem cells for 1-10 days in a stem cell culture medium configured by including a basal medium at 60.0–90.0 vol% and physiological saline at 10.0–40.0 vol%, the culture medium being further supplemented with 1.0–100.0 ng / mL of EGF, 0.2–20.0 ng / mL of FGF-2, 0.2–20.0 ng / mL of PDGF, and 0.5–8.0 mM of magnesium ascorbyl phosphate; b) thereafter, precipitating protein components from the culture solution obtained in step a) using an organic solvent; and c) isolating the precipitate portion and dissolving the same in a physiological saline or a basal medium to obtain a medium comprising a cell-produced protein (CPPs) composition, and seeding, using the medium, the mesenchymal stem cells at a density of 3×103 to 2×104 cells / mL and culturing for 1-3 days, thereby inducing differentiation of the mesenchymal stem cells into nerve 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.

Provided are a method of producing a human pluripotent stem cell-derived Leydig-like cell (Leydig-like cells) capable of secreting testosterone stably and persistently, and a human pluripotent stem cell-derived Leydig-like cell. Also provided is a human pluripotent stem cell-derived Leydig-like cell that enables long-term maintenance of a Leydig-like cell secreting a sufficient amount of testosterone, and is improved in efficiency of differentiation induction. The method includes the steps of: culturing a human pluripotent stem cell under a culture condition with addition of one or a plurality of kinds selected from the group consisting of: a WNT canonical pathway activator; bone morphogenetic protein 4 (BMP4); and vascular endothelial growth factor (VEGF); and forcedly expressing NR5A1, and is achieved by controlling the timing of addition and removal of cyclic adenosine monophosphate (cAMP), suspension culture, adhesion culture, and the like.

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 invention discloses the technical field of animal cell culture and cell culture meat, and provides a food-grade natural compound combination method for promoting efficient myogenic differentiation in vitro. According to the method disclosed by the invention, a combination of xanthophyll and arachidic acid is added into a basic myogenic differentiation induction culture medium to form a myogenic differentiation promoting culture medium. According to the myogenic differentiation culture medium system, the myogenic differentiation efficiency of the myogenic cells can be improved from about 23% to 45% or above, the myogenic fusion efficiency can be improved from about 18% to 33% or above, the problems that the in-vitro differentiation efficiency of the myogenic cells is low, the content of key proteins such as myosin is insufficient and the like are effectively solved, and the myogenic differentiation culture medium system is suitable for large-scale popularization and application. And a feasible scheme is provided for large-scale muscle fiber production of cell culture meat.

The invention provides an umbilical cord mesenchymal stem cell adipogenic induced differentiation culture medium and a culture method, and belongs to the technical field of stem cell induced differentiation. Comprising a DMEM culture medium, fetal calf serum, dexamethasone, insulin, indometacin, IBMX, Zardaverine and glabridin. Zardaverine and glabridin are added into an umbilical cord mesenchymal stem cell adipogenesis induced differentiation culture medium according to a certain concentration, and the synergistic effect of Zardaverine, glabridin and other components can better induce the umbilical cord mesenchymal stem cells to differentiate into adipocytes, increase the number of lipid droplets, increase the volume of the lipid droplets, enhance the lipid storage capacity of the adipocytes, and improve the adipogenesis of the umbilical cord mesenchymal stem cells. The adipogenic differentiation induction period is shortened, and the problems of low adipogenic differentiation rate, long induction time, few and small lipid droplets, unstable adipogenic effect and the like in the three-dimensional differentiation identification of the umbilical cord mesenchymal stem cells are solved.

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 present invention provides a prediction device, a prediction method, and a kit for predicting a future differentiation level of a cultured cell before start of differentiation induction, based on a content of miRNA in an exosome in the cultured cell, and an automatic cell culture system. Such implementations can involve a measurement device that is configured to measure the miRNA content in the culture supernatant of the cultured cell.

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 invention discloses a method for detecting the influence of bilirubin stimulation on human umbilical cord mesenchymal stem cells, and relates to the technical field of experimental medicine. Comprising the following steps: S1, culturing cells; s2, preparing a bilirubin solution; s3, detecting the cell survival rate by a CCK-8 method; s4, adipogenic differentiation induction; s5, osteogenic differentiation induction; s6, determining the concentration of cell factors through an ELISA method; and S7, detecting the expression of the apoptotic protein by using Western blotting. The invention provides a detailed and clear experimental method which is used for detecting the influence of bilirubin stimulation on human umbilical cord mesenchymal stem cells. The influence of bilirubin on hUC-MSCs can be comprehensively evaluated through the steps of detecting the cell survival rate through a CCK-8 method, inducing adipogenesis and osteogenesis differentiation, determining the cell factor concentration through an ELISA method, detecting apoptosis protein expression through Western blotting and the like.

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 invention relates to application of dendrobium officinale tender shoots and / or extracts thereof, in particular to application of dendrobium officinale tender shoots and / or extracts thereof obtained through differentiation induction of dendrobium officinale stem cells in preparation of drugs for treating and / or preventing glomerular cell fibrosis and / or renal podocyte injury related diseases.