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 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 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 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 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 supplement for inducing differentiation and maturation of stem cells to insulin-secreting cells in vitro and its use in inducing differentiation of stem cells. The supplement can promote the differentiation of stem cells from various sources into insulin-secreting cells in vitro, including adult stem cells derived from pancreatic tissue, induced pluripotent stem cells (iPS cells) or stem cells derived from other tissues. The main components of the culture supplement of the present invention are glucagon-like peptide 1 or its analogue, nicotinamide, and 1,25‑(OH)2D3. The culture supplement can be used in conjunction with any basal medium to induce stem cells to differentiate into insulin-secreting cells with good efficiency and the maturity of differentiated cells. The culture supplement disclosed by the invention can be used to induce differentiation of stem cells into insulin-secreting cells, so as to prepare stem cell products that can replace the function of pancreatic islets.

The invention discloses a kit for verifying endodontium mesenchymal 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 endodontium mesenchymal stem cells have positively-expressed surface markers CD146 which are different from the surface markers of other mesenchymal stem cells; CD146 antibodies are added into the kit, and accordingly, accurate verification results can be acquired.

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.