36results about How to "Improve the efficiency of induced differentiation" patented technology

The invention belongs to the field of regenerative medicine of cardiac muscle tissues, and discloses application of ascorbic acid as an induction factor in inducting differentiation of iPS (induced pluripotent stem) cells into cardiac muscle cells and a differentiation-inducing culture medium containing ascorbic acid. The invention also discloses a method for inducing differentiation of iPS cells into cardiac muscle cells by using the differentiation-inducing culture medium, which comprises the following step: inducing an iPS cell derived embryoid body, which has grown in a suspension mode for 5-7 days, to differentiate by utilizing the differentiation-inducing culture medium, thereby obtaining cardiac muscle cells. By using ascorbic acid as the induction factor, the invention obviously enhances the differentiation efficiency of the iPS cells into the cardiac muscle cells, and the differentiation percentage can reach 50-60%; the differentiation-inducing culture medium does not have toxic or side effect on the cells; and in addition, the method provided by the invention is simple, has the advantage of low cost, can be operated in common laboratories, and is suitable for large-scale proceeding.

The invention belongs to the technical field of cell induced differentiation, and in particular relates to an inducing agent and a culture medium for directional differentiation of an adipose-derived stem cell. The inducing agent for transformation of an adipose-derived stem cell into a testosterone cell consists of icariin, retinoic acid and human chorionic gonadotropin. The inducing culture medium for transformation of the adipose-derived stem cell into the testosterone cell is prepared by a method as follows: each 1000ml of the inducing culture medium consists of 0.5-10umol of icariin, 1-10umol of retinoic acid and 20,000-80,000 units of human chorionic gonadotropin, as well as a human mesenchymal stem cell serum-free medium, and the culture medium is obtained through uniformly mixing, filtering and sterilizing. The inducing agent and the culture medium for transformation of the adipose-derived stem cell into the testosterone cell disclosed by the invention are free from cell transfection, so as to avoid risks of gene modification and cancer, and induced differentiation efficiency is high; the inducing agent and the culture medium develop characteristics of traditional Chinese medicine, and are capable of inducing transformation of the autologous adipose-derived stem cell into the testosterone cell, and are free from an ethics problem and high in safety.

The invention discloses culture solution for inducing mesenchymal stem cells to differentiate into glomerular mesangial cells and an inducing method. The culture solution comprises a human platelet-derived growth factor-BB, all-trans-retinoic acid, collagen-IV and a basic medium, wherein the basic medium is a low-sugar DMEM (Dulbecco's modified eagle medium) containing 2% of horse serum and 5ng / ml of bFGF (basic fibroblast growth factor). The inducing method includes the steps: preparing the inducing culture solution; preparing the mesenchymal stem cells; coating a culture plate by the collagen-IV with the concentration of 5ug / cm<2>; inoculating the mesenchymal stem cells with the cell density of 5*103 / cm<2> into the coated culture plate and adding the inducing culture solution; changing the solution once every three days and performing inducing culture for seven days. The mesenchymal stem cells are jointly induced to differentiate into the glomerular mesangial cells by the aid of the human platelet-derived growth factor-BB, the all-trans-retinoic acid and the collagen-IV, inducing period is short, inducing efficiency is high, the glomerular mesangial cells formed by differentiation are high in functional activity, and transplantation therapy of kidney diseases such as kidney failure is facilitated.

The invention discloses a tissue culture rapid propagation method for coix lacryma-jobi. The method comprises the following steps: selecting a coix lacryma-jobi explant, performing sterilization treatment on the coix lacryma-jobi explant, and placing the coix lacryma-jobi explant which is subjected to the sterilization treatment in a coix lacryma-jobi callus differentiation culture medium for culturing and differentiating; after coix lacryma-jobi calluses are cultured, strengthening coix lacryma-jobi callus subculture through a coix lacryma-jobi callus subculture medium; performing coix lacryma-jobi redifferentiation culture through a coix lacryma-jobi redifferentiation culture medium; culturing by adopting a coix lacryma-jobi seedling culture medium when the coix lacryma-jobi grows into a seedling through coix lacryma-jobi redifferentiation; after the coix lacryma-jobi grows into the seedling, performing coix lacryma-jobi rooting culture through a coix lacryma-jobi rooting culture medium. The tissue culture rapid propagation method for the coix lacryma-jobi disclosed by the invention is high in callus induction differentiation efficiency, and good in induction seedling effect; coix lacryma-jobi planting resources can be preserved, a genetic transformation system can be established, and the production of a virus-free tissue culture seedling can be realized; therefore, the tissue culture rapid propagation method for the coix lacryma-jobi is worthy of being popularized.

The invention discloses a method for inducing and differentiating precursor cells of a human skin source into corneal endothelial cells. The precursor cells of the human skin source are used and cultured together with the corneal endothelial cells B4G12, corneal endothelial cells theoretically approaching corneal endothelial cells of a normal person are successfully induced, the obtained corneal endothelial cells are applied to a corneal endothelial decompensation animal model, the corneal endothelium of an animal is successfully repaired, and the method has important clinical application prospects.

The invention provides a method for inducing differentiation of adipose derived stem cells into chondrocyte. The method comprises the steps of preparing adipose derived stem cell-sodium alginate suspension, preparing adipose derived stem cell-calcium alginate microbeads, and performing induced culture. According to the method for inducing differentiation of adipose derived stem cells into chondrocyte, an incomplete cartilage forming induced culture solution containing PRP and transforming growth factor-beta superfamily members (TGF-[beta]1, IGF-1 and BMP-6) is utilized, and PRP and transforming growth factor-beta superfamily members are in united use, so that synergistic reactions are achieved, Sox-9 gene expression can be raised, differentiation of induced adipose derived stem cells intohypertrophy phenotypes is restrained, cell proliferation is promoted, and induced differentiation efficiency is improved. The proliferation vitality of the chondrocyte obtained through induced differentiation for 13-15 days by the method disclosed by the invention is 3.4 times of that of a control group A, 2.2 times of that of an experimental group B, and 1.95 times of that of an experimental group C.

The present invention provides a cell culturing device that, when culturing cells using a cell culture bag in which a plurality of recesses is formed, releases cells or cell aggregates adhering to theinner surfaces of the recesses while preventing movement of cells between the recesses, thereby improving the proliferation / differentiation induction efficiency of cells or cell aggregates. The present invention is a cell culturing method that uses a cell culturing bag (1) that comprises a bag body (2) configured from an upper surface film (21) and a lower surface film (22) having a sealed periphery, and a port (3) attached to the bag body (2), a plurality of recesses (4) being formed in the lower surface film (22), and the method comprising: a closing-off step for discharging a culture medium (S) contained in the bag body (2) through the port (3), and closing off the plurality of recesses (4) using the upper surface film (21); and a releasing step for releasing, in some or all of the plurality of closed-off recesses (4), spheroids adhering to the inner surfaces of the recesses (4).

The invention discloses a kit for identifying mesenchymal stem cells. The kit comprises a flowing phenotypic detection reagent suit, cell fixing liquid, a general culture medium, an adipogenesis differentiation induction and detection reagent suit, an osteogenesis differentiation induction and detection reagent suit and a chondrogenesis differentiation induction and detection reagent suit. The conventional osteogenesis and adipogenesis induction differentiation generally need about 24 days, so that the time is longer, and the induction efficiency is lower. The kit provided by the invention has the advantages that the osteogenesis and adipogenesis induction differentiation time is shortened to be 18 days; the induction differentiation efficiency is improved; the induction differentiation time is shortened; a flowing detection antibody is provided, so that a faster and more accurate authentication result is obtained.

The invention discloses an induction method of megakaryocyte progenitor cells. The method includes: isolating and screening cord blood CD34+CD126- cells; isolating and culturing umbilical cord mesenchymal stem cells; co-culturing the cord blood CD34+CD126- cells with the umbilical cord mesenchymal stem cells, and conducting amplification and induced differentiation of megakaryocyte progenitor cells. The induction method of megakaryocyte progenitor cells provided by the invention performs isolation and screening of cord blood CD34+CD126- cells and adopts the way of co-culture with the umbilicalcord mesenchymal stem cells for induced differentiation of megakaryocyte progenitor cells, increases the induced differentiation efficiency of in vitro megakaryocyte progenitor cells, and also enhances the amplification efficiency.

The invention discloses a culture solution for inducing mesenchymal stem cells to differentiate into islet-like cells, and an inducing method and application of the culture solution. The culture solution comprises the following materials: niacinamide, Conophylline, cell growth factor, betacellulin and a base medium. The base medium contains 97% of high glucose DMEM (Dulbecco Modified Eagle Medium), 2% of B-27 and 1% of N-2. The inducing method comprises the following steps of: preparing an inducing culture solution; preparing the human mesenchymal stem cells; taking the human mesenchymal stem cells, inoculating the human mesenchymal stem cells into a six-hole ultralow absorption culture plate by 1.5-2*105cells / hole, adding 3ml inducing culture medium into each hole and carrying out suspended induction; and changing liquid at every 3 days, collecting cell supernatant at the ninth day, and storing the cell supernatant at the temperature of -20 DEG C. The culture solution disclosed by the invention has the advantages that the human mesenchymal stem cells are induced to differentiate into the islet-like cells by utilizing the combination of the niacinamide and the Conophylline, so that the inducing cycle is shortened, the suspension cells are beneficial to being clustered to form cell clusters similar to natural islets, further the induced differentiation efficiency is obviously increased and the clinical application risk is reduced; and the function of inducing the secretion of the cell insulin is obviously improved.

The invention discloses an autophagy-based stem cell myocardial cell induced differentiation method and an application thereof. The method comprises the following steps of step 1, preparing an autophagy inducer wherein, the autophagy inducer dissolves in DMSO to prepare mother liquor, the mother liquor is stored at 20 DEG C, a solvent DMSO with the concentration being 0.01% is served as a controlgroup, and the mother liquor is added once when liquor is changed; step 2, stem cell culture; step 3, a first stage for stem cell induction, wherein a myocardial differentiation inducer is added, andcomprises the autophagy inducer prepared in the step 1 and vitamin C; step 4, a second stage for stem cell induction, wherein after good embryoid bodies are obtained through suspension culture, embryoid bodies are transferred into a cell culture plate for adherent culture, the induced culture solution is the same as that in the first stage, the liquor is exchanged each day or every two days according to the cell growth condition, and as time goes on, spontaneous rhythmic jump of the embryoid bodies is observed. The method improves the induced differentiation efficiency and is suitable for popularization and application.

The invention provides a method for controlling an incubator to be applicable to induction and amplification of immune cells. The method comprises the steps: a controller starts an oxygen source and acarbon dioxide source for supplying oxygen and carbon dioxide into a culturing space; the controller starts an oxygen concentration sensor and a carbon dioxide concentration sensor and acquires dataincluding oxygen concentration and carbon dioxide concentration in the space.

The invention provides an induced proliferation method of an immune cell. The method comprises the steps of: (1) inoculating a mononuclear cell into a bioreactor for cultivate for a day under an aseptic condition in 2-7% of CO2 and 8-10% of O2 at 37 DEG C, (2) continuously culturing the mononuclear cell cultured in step (1) for 14-21 days to form the immune cell, wherein an anti-human CD61 antibody is coated in advance in the bioreactor; the bioreactor comprises a first culture medium; the first culture medium is a T cell proliferation culture medium comprising 10% autologous plasma, 1000IU / mlIL-2 and 0.01KE / ml sapylin; the continuous culture from day 1 to day 8 in step (2) is performed in a second culture medium at 37 DEG C under the aseptic condition in 2-7% of CO2 and 8-10% of O2; thesecond culture medium is a T cell proliferation culture medium comprising 10% autologous plasma and 1000IU / ml IL-2, or a T cell proliferation culture medium comprising 1000IU / ml IL-2; the continuous culture from day 9 to day 21 is performed in a third culture medium at 37 DEG C under the aseptic condition in 12-18% of O2 and 2-7% of CO2; and the third culture medium is a SuperCulture TM L500 culture medium comprising 1000IU / ml IL-2.

The invention discloses a serum-free induction medium and an induction method for differentiating an iPS (induced pluripotent stem) cell into an endodermal progenitor cell. The serum-free induction medium comprises a first-stage medium, a second-stage medium and a third-stage medium. According to the serum-free induction medium and the induction method for differentiating the iPS cell into the endodermal progenitor cell, the iPS cell can be directionally induced to be differentiated into the endodermal progenitor cell, thereby providing a technical support for each iPS cell induced to be differentiated into an adult. In addition, no serum is used by the serum-free induction medium and the induction method, so that risks brought by animal-derived pathogens are effectively avoided, and the clinical application safety is improved.

The invention belongs to the field of biomedicine, and relates to an inducer for inducing mesenchymal stem cells to differentiate into islet cells. The invention discloses the inducer for inducing mesenchymal stem cells to be differentiated into islet cells. The inducer is prepared from the following components: GLP-1, parathyroid hormone, acetaminophen, rapamycin, icariin, trametinib, EPO and VEGF. According to the inducer for inducing and differentiating the mesenchymal stem cells into the islet cells, all the components are safe and non-toxic, the number of steps needed by induced differentiation is small, the time is short, and the induction efficiency is high.

The invention provides an umbilical cord Wharton's jelly mesenchymal stem cell osteogenic directional differentiation method and relates to the fields of stem cells and regenerative medicines. The method comprises the following steps of tissue digestion, primary culture, subculture, induction culture, etc.; in the tissue digestion step, tissue digestion liquid is used for tissue digestion, whereinthe tissue digestion liquid comprises collagenase I, DNA enzyme and Tryple; and in the induction culture step, an osteoblast differentiation induction culture medium is used for the induction culture, and the osteoblast differentiation induction culture medium comprises an alpha-MEM base bottom solution, a serum substitute, dexamethasone, beta-glycerophosphate, antibiotics, cytokine IL-beta, ascorbic acid and isoflavone. The method can obviously improve induction differentiation efficiency of umbilical cord mesenchymal stem cell osteoblasts.

The present invention discloses a method for resuscitating mononuclear cells and inducing into multi-cytokine-induced killer cells. The method comprises: placing cryopreserved mononuclear cells in a water bath with a temperature of 37-42 DEG C, and thawing to obtain thawed mononuclear cells; mixing the thawed mononuclear cells and a freezing resuscitation liquid for multi-cytokine-induced killer cells to obtain a resuscitation cell mixing liquid, wherein the freezing resuscitation liquid contains albumin and dextran; carrying out centrifugation treatment on the resuscitation cell mixing liquidto obtain resuscitated cells; and carrying out directional induction treatment on the resuscitated cells to obtain the induced multi-cytokine-induced killer cells. According to the present invention,the multi-cytokine-induced killer cells obtained through the induction and amplification of the mononuclear cells obtained through the resuscitating have advantages of rapid proliferation, high induction differentiation efficiency and good cell viability. .

The invention provides a culture medium for inducing differentiation of dental pulp stem cells into adipoblasts, a kit containing the culture medium and application of the kit. The culture medium comprises a basic culture medium, insulin, a platelet lysis buffer, indometacin, dexamethasone and 3-isobutyl-1-methylxanthine. The components contained in the culture medium have a good synergistic effect, so induced differentiation from the dental pulp stem cells to the adipocytes can be achieved, the adipogenic directional differentiation specificity of the culture medium is high, the specificity of the culture medium is high, time needed for the adipogenic induced differentiation of the dental pulp stem cells can be greatly shortened, and induced differentiation efficiency is improved. Meanwhile, the culture medium is applied to the kit for inducing the differentiation of the dental pulp stem cells into the adipoblasts. The kit is used for inducing the dental pulp stem cells, and stable and efficient induced differentiation of the dental pulp stem cells into the adipoblasts can be achieved.

The invention discloses a culture additive for in-vitro induction of stem cells to insulin-secretion cells differentiation maturation and a purpose of the culture additive in stem cell induction differentiation. The additive can promote in-vitro differentiation of stem cells with various sources to insulin-secretion cells, which comprises pancreas tissue-derived adult stem cells, induced pluripotent stem cells (iPS cells), or other tissue-derived stem cells. The culture additive comprises the main components of glucagon-like peptide1 or its analogue, niacinamide, and 1,25-(OH)2D3. The cultureadditive can be matched with any basic medium for usage, and the efficiency for induction of stem cell differentiation to the insulin-secretion cells and the cell maturity after differentiation are good. The culture additive can be used for induction differentiation of the stem cell to the insulin-secretion cells, so that the stem cell products capable of substituting pancreas islet function can be prepared.

The invention provides a culture medium and a method for inducing airway basal stem cells to differentiate into bronchoalveolar stem cells, and relates to the technical field of stem cell induced differentiation. The culture medium comprises a primary induced differentiation culture medium and a secondary induced differentiation culture medium; the primary induced differentiation culture medium comprises the following components: a fibroblast growth factor 10, a keratinocyte growth factor and a glycogen synthase kinase-3 inhibitor; the secondary induced differentiation culture medium is prepared from the following components: a fibroblast growth factor 10, a keratinocyte growth factor, a keratinocyte growth factor, a gamma secretase inhibitor and a phosphodiesterase inhibitor. According to the method disclosed by the invention, the BC is subjected to induction culture by adopting the induction culture medium and can be efficiently converted into the BASC. According to the culture medium and the culture method, autologous airway basal stem cells can be successfully induced into bronchoalveolar stem cells, and the induction conversion rate is high.

The invention discloses a tissue culture rapid propagation method for coix lacryma-jobi. The method comprises the following steps: selecting a coix lacryma-jobi explant, performing sterilization treatment on the coix lacryma-jobi explant, and placing the coix lacryma-jobi explant which is subjected to the sterilization treatment in a coix lacryma-jobi callus differentiation culture medium for culturing and differentiating; after coix lacryma-jobi calluses are cultured, strengthening coix lacryma-jobi callus subculture through a coix lacryma-jobi callus subculture medium; performing coix lacryma-jobi redifferentiation culture through a coix lacryma-jobi redifferentiation culture medium; culturing by adopting a coix lacryma-jobi seedling culture medium when the coix lacryma-jobi grows into a seedling through coix lacryma-jobi redifferentiation; after the coix lacryma-jobi grows into the seedling, performing coix lacryma-jobi rooting culture through a coix lacryma-jobi rooting culture medium. The tissue culture rapid propagation method for the coix lacryma-jobi disclosed by the invention is high in callus induction differentiation efficiency, and good in induction seedling effect; coix lacryma-jobi planting resources can be preserved, a genetic transformation system can be established, and the production of a virus-free tissue culture seedling can be realized; therefore, the tissue culture rapid propagation method for the coix lacryma-jobi is worthy of being popularized.

The invention relates to a method for inducing mesenchymal stem cells to differentiate into islet-like cells, and aims at providing the method for differentiating the mesenchymal stem cells coming different sources into the islet-like cells to obtain a large number of the islet-like cells. In order to achieve the aim, the method particularly comprises the following steps: 1) carrying out separation and primary culture on the mesenchymal stem cells of umbilical cord, placenta or fat sources; 2) preparing a differentiation medium: preparing BTCM, cell differentiation induced liquid and SCM; 3) inducing the mesenchymal stem cells to differentiate into the islet-like cells in the third stage; 4) obtaining identification of the islet-like cells. The islet-like cells obtained through the method are high in maturity, and less number of the cells can reach a fast hypoglycemic effect, so that the method for inducing the mesenchymal stem cells to differentiate into the islet-like cells has wide application prospects and is applied to the field of biomedicine.

The invention belongs to the technical field of induction and differentiation of stem cells, and in particular relates to an inducer of directed differentiation of mesenchymal stem cells. A mesenchymal stem cell epidermal differentiation inducer, which is composed of KGF, BMP‑4, PDGF‑AB, VEGF, IL‑2, and Forskolin. After adding complete medium, the concentration of each component is KGF 20~30μg / L , BMP-4 5-10 μg / L, PDGF-AB 5-10 μg / L, VEGF 10-20 μg / L, IL-2 5-10 μg / L, Forskolin 1-2 mg / L. The mesenchymal stem cell epidermal differentiation inducer of the present invention uses cytokines combined with small molecule compounds to efficiently induce mesenchymal stem cells to differentiate into epidermal cells, greatly improves the differentiation efficiency, and has strong cell activity after induction without cell transfection. Therefore, there is no genetic change and risk of cancer, no ethical issues, and high safety.

The invention discloses a method for inducing differentiation of human skin-derived precursor cells into corneal endothelial-like cells. The present invention successfully induces corneal endothelial-like cells that are theoretically close to normal human corneal endothelial cells by using human skin-derived precursor cells through co-culture with corneal endothelial cells B4G12, and applies the obtained corneal endothelial-like cells to corneal endothelial cells The decompensated animal model successfully repairs the corneal endothelial layer of animals, which has important clinical application prospects.

The invention discloses a kit for verifying adipose-derived stem cells. The kit comprises a flow phenotype detection reagent set, cell fixation liquid, an ordinary culture medium, an adipogenesis induction and detection reagent set, an osteogenesis induction and detection reagent set and a chondrogenesis induction and detection reagent set. Usually, conventional osteogenesis induction and adipogenesis induction need about 24 days, consumed time is long, and induction efficiency is low; by the kit, the time for osteogenesis induction and adipogenesis induction is shortened to 18 days, induction efficiency is improved, and induction time is shortened; conventional flow detection mainly aims at universal surface markers of mesenchymal stem cells, but the adipose-derived stem cells have positively-expressed surface markers CD49d and negatively-pressed CD106 which are different from the surface markers of other mesenchymal stem cells; CD49d antibodies and CD106 antibodies are added into the kit, and accordingly, accurate verification results can be acquired.

The invention discloses a serum-free induction medium and an induction method for differentiating an iPS (induced pluripotent stem) cell into an endodermal progenitor cell. The serum-free induction medium comprises a first-stage medium, a second-stage medium and a third-stage medium. According to the serum-free induction medium and the induction method for differentiating the iPS cell into the endodermal progenitor cell, the iPS cell can be directionally induced to be differentiated into the endodermal progenitor cell, thereby providing a technical support for each iPS cell induced to be differentiated into an adult. In addition, no serum is used by the serum-free induction medium and the induction method, so that risks brought by animal-derived pathogens are effectively avoided, and the clinical application safety is improved.

The invention provides a culture medium and a method for promoting induced differentiation of hematopoietic stem cells into megakaryocyte cells. The culture medium comprises a basic culture medium; a thrombopoietin; recombinant human stem cell factors; Interleukin-3; Interleukin-6; a poloxamer 188; and an aromatic hydrocarbon receptor antagonist. The culture medium of the invention can effectively promote induced differentiation of hematopoietic stem cells to megakaryocyte cells, improves the differentiation efficiency and the yield and activity of megakaryocyte cells, and has wide application prospects.

The invention discloses a culture medium for induced differentiation of CD34+ umbilical cord blood mononuclear cells. The culture medium consists of a basal culture medium and a composite additive; the composite additive consists of following components: 40-120 ng / ml of SCF; 4-8 ng / ml of IL3; 3-5 IU of Epo; 1-2 [mu]M of hydrocortisone or dexamethasone; 30-35 ng / ml of Flt3L; and 30-40 ng / ml of IGF-1. The invention further discloses a culture method for induced differentiation of the CD34+ umbilical cord blood mononuclear cells. The culture method comprises the following steps: (1) inoculating CD34+ umbilical cord blood mononuclear cells into a first-stage culture medium for culture; (2) from the seventh day, adding a second-stage culture medium for culture; (3) from the eleventh day, addinga third-stage culture medium for culture; and performing culture to the eighteenth day to obtain erythroid specific cells. According to the culture medium and the culture method, the induced differentiation efficiency of the CD34+ umbilical cord blood mononuclear cells can be remarkably improved, the differentiation and amplification time of the CD34+ umbilical cord blood mononuclear cells is shortened, and the erythroid specific cells are rapidly and efficiently obtained.