38 results about "Mesenchymal stem cell differentiation" patented technology

The invention discloses a method for transdifferentiating tail fin cells of erythroculter ilishaeformis into fat and muscle cells and application, the method is based on 5-Azacytidine (azacytidine) to promote mesenchymal stem cells to be differentiated into muscle cells, and other small molecule compounds are added on the basis of adding the 5-Azacytidine to transdifferentiate the tail fin cells of erythroculter ilishaeformis into muscles. The method is realized through an in-vitro cell reprogramming technology, fish individuals do not need to be sacrificed, and the destructive influence of traditional muscle or fat sampling on fish bodies is avoided; a single-step induction system does not need multi-factor sequential treatment, a high-purity target cell population can be obtained only in 5-7 days at least, the standardization degree of the operation process is high, the stability among batches is excellent, specific cells can be continuously and stably produced, and a safe and reliable technical basis is provided for sustainable cell culture fish meat production.

The invention relates to the technical field of stem cells, in particular to a serum-free induction medium with definite components for promoting and inducing mesenchymal stem cells to be differentiated into dermal papilla cells and an induction method. The small molecule compound composition is prepared from all-transretinoic acid, calomel, CHIR-99021 and minoxidil according to the molar ratio of (0.2 to 5): (2 to 50): (0.2 to 10): (0.2 to 10). According to the scheme, a plurality of small molecule compounds are adopted and have a synergistic effect, so that differentiation and function improvement of dermal papilla cells are promoted, and transformation of mesenchymal stem cells to the dermal papilla cells is promoted. The former three promote dermal papilla cell differentiation, and the latter promotes dermal papilla cell proliferation and inhibits apoptosis. According to the technical scheme, the technical problems that cell factors need to be added in the process of forming dermal papilla cells from stem cells through induced differentiation, so that the process cost is high, and the clinical safety is not ideal can be solved, the induction process is accelerated, and the method has ideal popularization and application prospects.

The present invention discloses the use of Avn Anthramides C (Avn C) as an inducer of chondrocytic differentiation in Mesenchymal Stem Cells (MSCs), providing a promising approach for regenerative cartilage therapy. By employing Avn C treatment, MSCs exhibit enhanced chondrocytic differentiation and elevated expression of key specific chondrogenic extracellular matrix (ECM) factors, notably Collagen Type II (COL2A1). Furthermore, when Avn C is used in combination with Chondroitin Sulfate (CS), a synergistic effect is observed, leading to upregulation of the transcriptional activity state of SOX9, a key regulator of chondrogenesis. The disclosed invention shows significant improvements in mRNA gene expression (ACAN, LRP1, SOX9), cellular protein levels (COL2A1, SOX9, LRP1), and functional outcomes, as evidenced by the secretion of extracellular matrix proteins (FSTL-1, BGH3, CO1A1, CO1A2, and CO3A1).

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.

The invention relates to a method for differentiating mesenchymal stem cells into adipose cells, which comprises the following steps: (1) culturing early passage mesenchymal stem cells to a confluence degree close to 100%, and ensuring enough basic cell quantity; (2) replacing the basic culture medium with a fat induction culture medium, and performing induced differentiation for 14-21 days; (3) observing a saturated oil red O dyeing result; and (4) the method can be applied to differentiation of mesenchymal stem cells in animal cell culture meat into adipocytes to form lipids, and is suitable for differentiation of mesenchymal stem cells into adipocytes in stem cell treatment and health care.

This invention discloses a method for preparing a 3D-printed hydrogel scaffold for cartilage repair, comprising: providing methacrylamide chitosan; providing 2-([1,1-biphenyl]-4-ylcarbamoyl)benzoic acid@polylactic acid-glycolic acid copolymer microspheres; obtaining a 3D-printed bio-ink for cartilage repair using methacrylamide chitosan, 2-([1,1-biphenyl]-4-ylcarbamoyl)benzoic acid@polylactic acid-glycolic acid copolymer microspheres, recombinant collagen CF-1552, and lithium magnesium silicate as main raw materials; and obtaining a 3D-printed bio-ink for cartilage repair using the 3D-printed bio-ink for cartilage repair as a raw material to obtain a 3D-printed hydrogel scaffold for cartilage repair. This 3D-printed hydrogel scaffold has advantages such as antibacterial properties and the ability to promote the differentiation of human bone marrow mesenchymal stem cells (hBMSCs) into chondrocytes, and can be used for cartilage repair, potentially bringing new curative opportunities for cartilage injury diseases.

The application discloses a culture solution for indirectly inducing human induced pluripotent stem cells into mesenchymal stem cells, which is composed of a neuroectoderm induction culture solution, a mesenchymal stem cell differentiation culture solution and a mesenchymal stem cell maintenance culture solution. The neuroectoderm induction culture solution is composed of a Neurobasal culture medium, a DMEM / F12 culture medium, an N2 additive, a B27 additive, a GSK3 inhibitor and a TGF-beta inhibitor. The application further discloses a method for indirectly inducing human induced pluripotent stem cells into mesenchymal stem cells, which comprises the following steps: taking the induced pluripotent stem cells, culturing the induced pluripotent stem cells in the neuroectoderm induction culture solution for 6 days, then culturing the induced pluripotent stem cells in the mesenchymal stem cell differentiation culture solution for 7 days, and then culturing the induced pluripotent stem cells in the mesenchymal stem cell maintenance culture solution for 3-5 generations. The method can direct the induced pluripotent stem cells to differentiate into mesenchymal stem cells in a short time, and greatly shortens the culture time.

The present invention provides a method for promoting the differentiation of mesenchymal stem cells (MSCs) into oligodendrocyte progenitor cells, which includes primary culturing the isolated MSCs, followed by secondary passage culturing, then treating them with a gene expression vector containing the "key gene for oligodendrocyte progenitor cell differentiation", and further performing differentiation culture using the "oligodendrocyte progenitor cell differentiation medium". Through specific cell differentiation treatment and culture processes, the present invention uses a cell culture matrix treated with recombinant human laminin to culture MSCs and promote their differentiation, so as to obtain oligodendrocyte progenitor cells derived from MSCs in a relatively short time. The present invention further provides a method for efficiently harvesting oligodendrocyte progenitor cells. Specifically, through a rate-limiting shaking method, oligodendrocyte progenitor cells are harvested and allowed to further proliferate, and a relatively large amount of oligodendrocyte progenitor cells with high purity can be harvested in a short time for the production of cell therapy products.

The invention discloses a preparation method and application of a leukocyte-inducing extract. The preparation method comprises the following steps: acquiring mesenchymal stem cells; inducing the mesenchymal stem cells to differentiate to generate dendritic cells; splitting the dendritic cells, and separating to obtain a cell extract; the differentiation comprises the steps of inducing the mesenchymal stem cells to differentiate into immature dendritic cells through the first induction composition and inducing the immature dendritic cells to mature through the second induction composition. The preparation method provided by the embodiment of the invention finds a new way, starts from the mesenchymal stem cells, and utilizes the characteristic of wide distribution of the mesenchymal stem cells to solve the problems of single source and difficult acquisition of leukocytes in the production process of the leukocyte extract; and moreover, the mesenchymal stem cells are differentiated, so that the situation that monocytes obtained by directly separating the albuginea layer are mixed into inflammatory cells is avoided.

To provide osteoblasts and compositions comprising the osteoblasts for treating bone diseases.SOLUTION: Disclosed is an osteoblast having both proliferation ability and differentiation ability, where the osteoblast has higher expression levels of connexin 43, RUNX2 and collagen type 1A compared with those of undifferentiated stem cells and mature osteocytes; higher expression levels of angiopoietin and alkaline phosphatase compared with those of undifferentiated stem cells; and lower expression levels of osterix, osteocalcin and osteopontin compared with those of mature osteocytes. Also, the osteoblast has a lower Ki-67 expression level compared with that of undifferentiated stem cells.SELECTED DRAWING: Figure 1

Provided is a method for inducing differentiation into nerve cells, wherein: a) mesenchymal stem cells are cultured for 1-10 days in a stem cell culture medium that is constituted by containing 60.0-90.0 vol% of a basal medium and 10.0-40.0 vol% of physiological saline and 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, a protein component in a culture solution that has been obtained by the culturing are precipitated with an organic solvent; and c) using a medium that contains a cell-produced protein (CPPs) composition obtained by isolating a precipitate portion and dissolving the precipitate portion in physiological saline or a basal medium, the mesenchymal stem cells seeded at a density of 3×104 to 2×105 cells / mL are cultured at 4-10°C for 8 hours to 2 days, thereby inducing differentiation of the mesenchymal stem cells into nerve cells.

The application provides a method for improving the content of cartilage glycosaminoglycan by using cationic polymer, and the method promotes the content of glycosaminoglycan in cartilage tissue to be obviously increased by treating cartilage with a preparation containing polybrene, creates a good microenvironment for cartilage repair, and promotes the cartilage to be better repaired and regenerated; the method can improve the content of glycosaminoglycan in the differentiated tissue of mesenchymal stem cells by culturing the mesenchymal stem cells in an induction medium containing polybrene, promotes the mesenchymal stem cells to be better differentiated into cartilage, and has a wide clinical application prospect.

The present invention is to provide a 3D cartilage organoid block prepared by differentiating mesenchymal stem cells into 3D spheroid cartilage tissues, a basic unit for the 3D cartilage spheroid block. The inventors found that both the amount of GAG matrix and the expression of the collagen type2 increased. Therefore, the method of this invention provides clinically applicable cartilage tissues by effectively enhancing the function of the cartilage differentiation constructs according to 2D culture. The 3D cartilage organoid block can be usefully applied to the area, such as, articular cartilage regeneration and plastic surgery, where cartilage tissues restoration is required.

The invention relates to the technical field of biomedicine, in particular to a mesenchymal stem cell differentiation method for promoting bone joint repair, which comprises the following steps: inoculating mesenchymal stem cells on a three-dimensional scaffold material to form a complex, applying an alternating electric field within a specific parameter range for stimulation, and combining with growth factor culture to realize directional differentiation. According to the method, by regulating and controlling the frequency, the voltage amplitude and the action time of the alternating electric field, the differentiation efficiency of the cells in the cartilage or osteogenesis direction is remarkably improved, and meanwhile the adaptability of the cells in the complex bone joint microenvironment is enhanced.

The invention relates to the technical field of biological cell culture, and the biological cell culture medium comprises a basic nutrient component, a signal regulation factor, a differentiation promoting factor and an auxiliary stable factor, and the unit of each component is mg / L according to the total volume of the culture medium; the basic nutrition component is prepared from the following substances: 200 to 800 L of glutamine, 150 to 600 L of serine, 300 to 1200 L of proline and 100 to 400 L of lysine hydrochloride. The invention relates to the technical field of biological cell culture. According to the mesenchymal stem cell induced chondrogenic differentiation culture medium and the preparation method thereof, by optimizing the composition of the culture medium, a specific activity enhancing component and a signal regulating component are selectively introduced, so that the efficiency of differentiating mesenchymal stem cells into chondrocytes is remarkably improved; an amino acid derivative in the activity enhancing component can effectively promote cell metabolism activity, and a growth factor simulant and a signal channel regulator in the signal regulating component synergistically act on a key signal channel in the cell differentiation process.

A method for mesenchymal stem cell differentiation from human pluripotent stem cells is provided, particularly including the step of adaptation of embryoid bodies from 3-dimensional culture to 2-dimensional culture. The method enables rapid mesoderm and mesenchymal stem cell transformation by transferring from 3-dimensional culture to 2-dimensional culture. Also, the method relates to obtaining mesenchymal stem cell, which can be used in clinical applications, with this method.

Provided are a differentiation-inducing culture medium for the transformation of adipose-derived mesenchymal stem cells into chondrocytes and the use thereof. A basal culture medium of the differentiation-inducing culture medium comprises a nutrient solution and an inducer. The nutrient solution comprises a blood substitute, a non-essential amino acid, β-mercaptoethanol, penicillin / streptomycin, dexamethasone, sodium pyruvate, ascorbic acid, sodium selenite, transferrin, recombinant human insulin, β-glycerophosphate and TGF-β1; and the inducer comprises cyanamide, delphinidin chloride and an extracellular matrix derived from adipose-derived mesenchymal stem cells. The components of the inducer, i.e. cyanamide and delphinidin chloride, can stimulate chondrocyte generation by means of up-regulating the expression of aggrecan and collagen II; and the component of the inducer, i.e. extracellular matrix (ADSC-ECM), can increase the adhesion between chondrocytes, enhance the signaling communication between chondrocytes, and promote the growth and metabolism of chondrocytes, thereby greatly improving the differentiation efficiency of adipose-derived mesenchymal stem cells into chondrocytes.

The invention relates to the technical field of biological culture, and particularly discloses a culture medium for improving synthesis of unsaturated fatty acid in cell culture fat, which is used for solving the problem that the content of unsaturated fatty acid in the culture fat is not satisfactory. The differentiation culture medium comprises the following components: a basic culture medium, a specific porcine adipose-derived mesenchymal stem cell differentiation induction factor and an unsaturated fatty acid additive. Wherein the specific induction factors comprise but are not limited to factors such as insulin, dexamethasone, indometacin, 3-isobutyl-1-methylxanthine, rosiglitazone and the like. The unsaturated fatty acid additive comprises alpha-linolenic acid, and by adding the substances into the culture medium, the differentiation efficiency of the porcine adipose-derived mesenchymal stem cells and the directional synthesis of the unsaturated fatty acid in the cells can be improved. According to the improved differential culture medium, cell differentiation and unsaturated fatty acid synthesis are effectively promoted by optimizing components and proportions, and the improved differential culture medium has an important application prospect.

The utility model belongs to the technical field of induction kits, and particularly relates to a mesenchymal stem cell differentiation induction kit which comprises a box body and a fixing component arranged on the outer side of the end part of the box body, the fixing assembly comprises a telescopic rod, a connecting plate and a sliding block, and the telescopic rod is installed on the inner side of the bottom end of the box body. By means of the fixing assembly, it can be guaranteed that the reagent tube is stable and immovable in the operation process, shaking or falling possibly occurring during manual operation is avoided, and therefore experiment accidents are reduced, when the reagent tube needs to be taken out, the fixing assembly can lift the reagent tube upwards to the proper height, an operator can grab the reagent tube easily and accurately, the experiment process is accelerated, and the experiment efficiency is improved. The time consumption is reduced, and the compact design of the box body is combined with the efficient fixing assembly, so that the limited space of a laboratory can be more effectively utilized.

The invention provides a preparation method of adipose-derived stem cells and application of the adipose-derived stem cells in neural restoration, and further, a specific differentiation-promoting peptide CF-8 for promoting differentiation of adipose-derived stem cells into neuron-like cells is screened from pilose antler polypeptides. The neuron-like cells generated by differentiation after the action of the differentiation promoting peptide have a good treatment effect on traumatic brain injury, and the application prospect is good.

The invention relates to the technical field of islet cell culture, and discloses a method for inducing mesenchymal stem cells to be differentiated into islet-like cells and application of the islet-like cells, and the method comprises the following steps: inoculating the mesenchymal stem cells into an umbilical cord-based tissue cell culture medium A for adherent multiplication culture; preparing the amplified mesenchymal stem cells into a cell suspension, and aggregating the cells into balls through centrifugation or limited culture; carrying out induced culture on the mesenchymal stem cell spheres by using a culture medium B; carrying out induced culture on the started cell balls by using a culture medium C; and carrying out induced culture on the differentiated cell balls by using a culture medium D to obtain the islet-like cells. Mesenchymal stem cells are prepared into cell spheres to enhance the intercellular interaction, and a culture medium B containing activin A and all-transretinoic acid, a culture medium C containing high glucose and nicotinamide and a culture medium D containing trichostatin A and Exendin-4 are used for staged induction to obtain the islet-like cells with the insulin secretion function.

The present application relates to the technical field of stem cells, and particularly relates to a serum-free induction medium with clear components for promoting differentiation of induced mesenchymal stem cells into dermal papilla cells and an induction method. A small molecule compound composition comprises all-trans retinoic acid, forskolin, CHIR-99021 and minoxidil in a molar ratio of 0.2-5:2-50:0.2-10:0.2-10. The present application adopts multiple small molecule compounds to promote differentiation and function improvement of dermal papilla cells and transformation of mesenchymal stem cells into dermal papilla cells. The first three promote differentiation of dermal papilla cells, and the last one promotes proliferation and inhibits apoptosis of dermal papilla cells. The present application can solve the technical problems of high process cost and unsatisfactory clinical safety caused by the need to add cytokines in the process of inducing differentiation of stem cells into dermal papilla cells, and speeds up the induction process, and has an ideal application prospect.

The present invention relates to a method for differentiating mesenchymal stem cells into osteoblasts, a cell therapy agent for treating bone diseases comprising osteoblasts differentiated by the above method, or a method for preparing the same. Furthermore, the present invention relates to a method for treating bone diseases, comprising administering osteoblasts obtained by the above method to a patient with a bone disease. The differentiation method of the present invention can stably and rapidly differentiate mesenchymal stem cells into osteoblasts. The above-mentioned differentiated osteoblasts have excellent angiogenesis ability and excellent bone formation ability. Therefore, the method of differentiating stem cells into osteoblasts of the present invention or the osteoblasts obtained by the above method can be effectively used as a cell therapy agent or treatment method related to bone diseases.

The invention discloses an electroactive bone repair membrane material with a bone-imitating unit structure as well as a preparation method and application of the electroactive bone repair membrane material. Research shows that the bioactivity of the bone repair membrane material can be improved by forming a bone-imitating unit structure on the electroactive bone repair membrane material. The electroactive bone repair membrane material with the imitated bone unit structure can effectively promote mesenchymal stem cell differentiation and up-regulate expression of tissue repair related genes, and can promote vascularized bone regeneration in vivo through an osteogenesis-vascularization synergistic effect.

The application provides a preparation method of fat stem cells and the use thereof in nerve repair, and further, a specific differentiation-promoting peptide CF-8 for promoting differentiation of fat mesenchymal stem cells into nerve-like cells is screened from a deer antler polypeptide, wherein the nerve-like cells differentiated after the action of the differentiation-promoting peptide have a good therapeutic effect on treating traumatic brain injury, and have a good application prospect.

The present disclosure belongs to the field of biological medicines, and relates to an inducer for inducing differentiation of mesenchymal stem cells into estradiol-secreting cells. The inducer for inducing differentiation of mesenchymal stem cells into estradiol-secreting cells uses human mesenchymal stem cell serum-free culture medium as a substrate and comprises the following components in mass concentration ratios: 20-60 mg / L of bone morphogenetic protein-4, 20-60 mg / L of bone morphogenetic protein-7, 2-8 mg / L of retinoic acid, 2-8 mg / L of resveratrol, 2-8 mg / L of icariin, 2-8 μg / L of benzamide, 2-8 μg / L of chloroplatinic acid hexahydrate, 2-8 μg / L of ethanolamine, 2-10 μg / L of erythropoietin and 2-10 μg / L of vascular endothelial growth factor. The inducer for inducing differentiation of mesenchymal stem cells into estradiol-secreting cells provided by the present disclosure has a high induction efficiency.

The invention discloses a culture medium for promoting differentiation of mesenchymal stem cells and a culture method, and belongs to the technical field of biology. The culture medium comprises a basic culture medium, beta-nicotinamide mononucleotide (0.3 to 6.0 mM), atractylenolide II (3 to 10 nM), BMP-4 (1 to 5 ng / mL) and glutathione (50 to 100 [mu] g / mL). Osteogenic differentiation related signal pathways are activated through the synergistic effect of multiple factors, and the differentiation efficiency is remarkably improved to 85% or above. Experiments show that the culture medium has a remarkable effect in bone marrow-derived mesenchymal stem cells, the calcification area is increased by 19.9% compared with that of a basic culture medium, the problems of low efficiency and high cost of a single inducible factor in the prior art are solved, and the culture medium is suitable for bone tissue engineering and clinical bone defect repair and has wide clinical application value.

A method for preparing mesenchymal stem cells (MSCs) is provided. The method includes providing expanded potential stem cells or a first cell culture including the expanded potential stem cells; culturing the expanded potential stem cells or the first cell culture in a trophoblast stem cell (TSC) differentiation medium to obtain trophoblast stem cells or a second cell culture including the trophoblast stem cells; culturing the trophoblast stem cells or the second cell culture in a mesenchymal stem cell (MSC) differentiation medium to obtain mesenchymal stem cells or a third cell culture including the mesenchymal stem cells; and passaging the mesenchymal stem cells or the third cell culture in an MSC expansion medium while maintaining characteristics of the mesenchymal stem cells.