151results about How to "High differentiation efficiency" patented technology

The invention discloses a method for inducing megakaryoblast and megakaryocyte in vitro and a special culture medium thereof. The special culture medium is a serum free medium containing 50ng/mL of SCF, 50ng/mL of TPO, 20ng/mL of IL-3 and 50ng/mL of IL-6. The method comprises the following steps of: 1) separating single karyocyte from umbilical cord blood by using the conventional Ficoll density gradient centrifugation method, wherein red blood cells in the umbilical cord blood are settled by 6 percent hydroxyethyl starch; and 2) culturing the single karyocyte in vitro for 4 to 14 days by using the special culture medium at 37 DEG C in the presence of 5 percent CO2 to obtain a mixture of the megakaryoblast and the megakaryocyte. The invention provides a source for supplementing the megakaryoblast and the megakaryocyte, and has the advantages of simple operation, low cost, high differentiation efficiency and the like. The method is about to play an important role in the field of medicaments, and has wide application prospect.

The invention discloses a method for differentiating human embryonic stem cells into hepatic cells by in vitro inducement, and a special culture medium thereof. The in vitro inducement method comprises the following steps: 1). human embryonic stem cells are suspended by a differential medium I; cell suspension is filled in a low absorbability germ culture dish, and is cultured for one to three days in 5 percent CO2 at 37 DEG C to form an embryoid body which is cultured for two to four days in a differentiating culture medium under the same conditions and is differentiated into sizing endoblast cells; 2). the sizing endoblast cells differentiated by the step 1 are inoculated to human fetal liver stroma cell culture layers of expressing basic fibroblast growth factors and are added with a differentiating culture medium III; after the sizing endoblast cells are cultured for four to six days in 5 percent CO2 at 37 DEG C, the culture medium III is replaced by a differentiating culture medium IV; and the sizing endoblast cells are continuously cultured for five to twenty days under the same conditions so as to obtain liver cells. The invention has the advantages of simple operation, low cost, high differentiating efficiency, and the like, exerts important action in medical fields, and has wide application prospects.

The invention discloses a method for breeding chamomile. The method disclosed by the invention comprises the following steps: (1) inoculating chamomile seeds onto a seed germination culture medium to carry out sterile seedling cultivation so as to produce sterile seedlings; (2) taking hypocotyls of the sterile seedlings as explants, inoculating the explants onto a callus induction culture medium to carry out callus induction culture so as to generate calluses by virtue of induction; (3) inoculating the calluses generated by induction onto an adventitious bud differentiation culture medium to carry out adventitious bud induction differentiation culture so as to obtain adventitious buds; (4) inoculating the adventitious buds onto a rooting culture medium to carry out rooting culture and culturing adventitious roots by virtue of induction so as to obtain rooted seedlings; and (5) hardening and transplanting the rooted seedlings so as to finally obtain the chamomile. In a regeneration system established by the method disclosed by the invention, callus inductivity reaches up to 86.63%, differentiation rate of the adventitious buds reaches up to 25.5%, rooting rate reaches up to 100%, and transplanting survival rate reaches up to 100%, thus a large number of excellent chamomile test-tube plantlets can be obtained in short term so as to realize large-scale factory production.

The invention relates to the technical field of regenerative medicine and discloses a method for adjusting and controlling in vitro three-dimensional directed differentiation of stem cells. The method includes following steps: embedding Stem cells into calcium alginate micro gel beads containing chondroitin sulfate, adding an inductive differentiation culture liquid, adjusting and controlling the in vitro directed differentiation of the stem cells by changing a mass ratio between the sodium alginate and the chondroitin sulfate during the preparation of the micro gel beads, dissolving the calcium alginate micro gel beads in a sodium citrate solution after the differentiation process being finished, and then obtaining pure differentiation cells. The method is simple in operation and low in cost, the calcium alginate micro gel beads containing chondroitin sulfate can simulate an in vivo three-dimensional extracellular matrix so that a three-dimensional directed differentiating microenvironment which approximates an in vivo microenvironment is supplied for the stem cells, thereby improving directed differentiation efficiency and a biological function of the stem cells. The method plays an important role in applications of regenerative medicine.

The invention relates to a method for inducing a human pluripotent stem cell to differentiate into an RPE cell. According to the method, an inhibitor is used for inducing the human pluripotent stem cell to differentiate into a neural precursor; after Shh, IGF1 and EGF growth factors are utilized to activate ERK 1/2, JNK, Sonic hedgehog signal routes, the cell is presented as the shape of neuroepithelium, Activen A and Chir99021 are added to activate TGFbeta and Wnt signal routes respectively to induce a neuro-epithelial cell into a retinal progenitor cell; adherent culture is conducted on theretinal progenitor cell to obtain an RPE cell at an underdeveloped state. The invention further studies the field of applying the underdeveloped RPE cell to treating retinal degenerative diseases. Compared with the prior art, the method firstly makes three stages of differentiating the human pluripotent stem cell to the underdeveloped RPE cell and related small molecular substances and growth factors used therein clear. Compared with a matured RPE, the underdeveloped RPE obtained through differentiation can treat the retinal degenerative diseases more effectively after being transplanted.

The invention relates to the technical field of stem cells, and in particular to a human mesenchymal stem cell osteogenic induction differential medium and a preparation method. The human mesenchymal stem cell osteogenic induction differential medium comprises an alpha-MEM/HG-DMEM medium as well as the following components in concentration: 5-50% by volume of fetal calf serum, 10-100mu M of ascorbic acid, 5-50mM of phosphoglycerol, 50-500nM of dexamethasone, 0.1-10nM of an insulin-like growth factor-1 and an 0.5-50nM of insulin-like growth factor-2. By adopting the medium, human mesenchymal stem cell osteogenic induction differential signal activation is intensified through the insulin-like growth factor-1 and the insulin-like growth factor-2, osteoblast proliferation is promoted, then the human mesenchymal stem cell osteogenic induction differential efficiency and specificity are improved, the differential time is shortened, the induction efficiency is improved, meanwhile the preparation method is simple and convenient and convenient to use, and stable and efficient human mesenchymal stem cell osteogenic induction differential can be achieved.

The invention is directed to methods for promoting differentiation of stem cells to hematopoietic cell lineages. The invention is further directed to increasing the differentiation efficiency of hematopoietic stem/progenitor cells. Such methods utilize novel compositions, including but not limited to, Amnion-derived Multipotent Progenitor cells (herein referred to as AMP cells) and conditioned media derived therefrom (herein referred to as Amnion-derived Cellular Cytokine Solution or ACCS), each alone or in combination with each other or other agents.

The invention provides a method for rapid propagation of pinellia ternate by tissue culture. The method comprises the steps as follows: (1) preparation of culture media; (2) selection of explants; (3)obtaining of callus by induction; (4) multiplication culture of callus; (5) differentiation culture of the callus; (6) acceleration culture; (7) strong seedling culture; (8) acclimatization and transplantation. By use of the special culture media and liquid culture of the callus for 10 d, the volume and weight are increased by 300%, and the quality of the callus is better; when green bud points appear in solid culture media, the callus is transferred to liquid culture media again, the callus differentiates after 10 d of culture, a large number of root systems and bud points appear, and the volume and weight are obviously increased; then, the callus is transferred to the solid culture media, root systems continuously grow, the bud points continuously develop to form a large number of leaves, complete pinellia ternate plants can be formed after 15 d, the propagation coefficient reaches 20 or above, the rooting rate reaches 95% or above, and the transplantation survival rate is 99% or above.

The invention discloses a neural stem cell induction differentiation medium and a neural stem cell induction differentiation method. Specifically, the neural stem cell induction differentiation mediumcomprises a basic medium and additives. The basic medium is a DMEM/F12 medium. The additives include 1-5mM glutamine, a 2-8microM GSK-3 inhibitor, a 2-10microM BMP inhibitor, a 2-10microM ALK inhibitor, 0.5%-2% of a cell culture additive and 0.5%-2% of a double-antibody. The neural stem cell induction differentiation medium provided by the invention has the characteristics of clear and simple components, high differentiation efficiency, short differentiation time, no animal-derived component, no need for animal-derived cells to serve as the feeder layer during differentiation, no need to formEB, and no need for repeated replacement of the culture solution components in differentiation, and can acquire a large number of high purity neural stem cells within a short time.

The invention belongs to the technical field of cytobiology and bone tissue engineering, relates to a compound used for treating osteoporosis, and especially relates to an application of raloxifene in mesenchymal stem cell in-vitro osteoblast differentiation. According to the invention, raloxifene is added during a mesenchymal stem cell in-vitro osteoblast differentiation culturing process. With the application, mRNA and protein level expressions of osteoblast differentiation marker genes ALP and BMP2, and calcium ion sedimentation can be increased; and mesenchymal stem cell differentiation towards osteoblast can be promoted. The invention relates to the application of raloxifene in mesenchymal stem cell in-vitro osteoblast differentiation, and in treatments of diseases such as fractures,bone defects, and bone wounds.

The invention discloses a culture solution for inducing multifunctional stem cells to differentiate into dopamine progenitor cells and a differentiation method. The culture solution comprises a DMEM/F12 culture medium and a Neurobaal culture medium in a volume ratio of 1: (0.9-1.3), and an N2 supplement, a B27 supplement, Lascorbic acid, Lglutamine, a GSK3 beta inhibitor, a TGF beta inhibitor, a BMP inhibitor, a fibroblast growth factor, a human leukemia inhibition factor hLIF, heparin Heparin and a nutritional factor required for growth of dopaminergic progenitor cells. The culture solution disclosed by the invention can be used for obtaining dopamine progenitor cells by taking induced multifunctional stem cells as raw materials through directional differentiation culture, can be used for direct transplantation or final differentiation into dopamine neurons, and can be used for clinical cell substitution transplantation treatment of PD, in-vitro disease modeling research, drug screening and other biomedical applications. The method has the advantages of being convenient to operate, short in culture time, high in yield, high in differentiation purity, free of animal-derived components in the whole link and the like.

The invention discloses a method for disintegrating human pluripotent stem cells into macrophages. The invention provides a reagent kit for disintegrating the pluripotent stem cells into the macrophages. The reagent kit comprises a complete set of culture fluid which can disintegrate the pluripotent stem cells into the macrophages. A monolayer cell culture method is adopted, firstly, hemutopoiesisstem cells are obtained through disintegrating, and then through further adding IL3 and M-CSF for stimulation, CD14 and CD163 positive macrophages are obtained. The method has the advantages that chemical components are definite, animal-source components do not exist, the safety of preparation cells is greatly increased, and the method has the characteristics of short in time consumption, high indisintegrating efficiency, low in cost and the like. Through the adoption of the preparation method provided by the invention, human macrophages can be produced in a large-scale manner, the quality is stable, the safety is high, and massive cell sources are provided for tissue project, medicine research and development and cell therapy.

The invention discloses a reprogramming method for efficient inducing of T cells into multipotent stem cells. The method comprises the following steps: S1, extracting monocyte from a blood sample, andadding the monocyte to an amplification culture system containing an activating agent for implementing selective T cell activating culture; S2, introducing an episomal vector which contains at leastone potential determinant into the T cells which are obtained in the step S1; S3, cultivating the T cells which are obtained in the step S2 and contain the episomal vector via a multipotent stem cellinducing medium, and performing inducing in a system free from a feeding layer, so that preprogrammed intermediate cells are obtained; and S4, after complete inducing, replacing the multipotent stem cell inducing medium in the step S3 with a multipotent stem cell medium for maintaining culture, so that cells that the expression of the potential determinant disappears and expression of endogenous multipotent genes, namely POU5F1, NANOG, TRA-1-60 and TRA-1-81, is activated are obtained, wherein the cells are induced multipotent stem cells. The method provided by the invention has the beneficialeffect that the T-cell derived induced multipotent stem cells can be simply and conveniently prepared in a large scale.

The invention discloses a rapid and high-efficiency transgenic method for indica rice. The method comprises the following steps of: exposing indica rice mature seeds to Agrobacterium tumefaciens 5 to 15 days after callus induction, co-culturing, performing resistant callus screening for 10 to 20 days, performing differentiation culture for 10 to 20 days, performing rooting culture for 5 to 10 days, and performing resistance screening again. The transgenic plants of indica rice can be produced within 40 to 50 days, and the conversion efficiency reaches 20-30%. In the conversion process, the culture conditions are as follows: the temperature ranges from 30 DEG C to 33 DEG C and the light is continuously supplied with the intensity of 80 to 120 mu mole/m<-2>s<-1>, except that the co-culture stage is carried out at 25 DEG C in the dark. The method provided by the invention greatly shortens the in vitro culture time of calli, increases the differentiation efficiency, reduces the mutation frequency of somatic cells, simplifies the operation procedure and saves a large amount of labor and material resources.

The invention provides a culture medium for differentiating neural stem cells into oligodendrocyte progenitor cells. The culture medium does not contain exogenous factors, so that pollution of the exogenous factors is avoided, and the oligodendrocyte progenitor cells can be efficiently differentiated. The invention further provides a method for preparing the oligodendrocyte progenitor cells by using the culture medium. By use of the method provided by the invention, the differentiation efficiency of the oligodendrocyte progenitor cells is improved on the premise of increasing the yield of theoligodendrocyte progenitor cells.

The invention discloses a method for increasing the efficiency of inducing pluripotent stem cells to be differentiated into hematopoietic stem cells in vitro. The method comprises the steps of carrying out in-vitro co-culture on OP9 and iPSCs, and meanwhile, adding trophic factors such as SCF, BMP4, IL-3, IL-6 and TPO into a differential medium. By using the method, the double positive rate of CD34+ CD43+ is increased to 15.3%, the differentiation efficiency is remarkably increased, and therefore, iPSCs can be induced to be differentiated into hematopoietic stem cells through adding a positive regulating factor, and the differentiation efficiency can be conveniently and rapidly increased only through a cell factor adding method.

The invention relates to the field of cellular biology, in particular to a retinal pigment epithelial cell and a preparation method and application thereof. The retinal pigment epithelial cell expresses MITF and ZO-1. The preparation method of the retinal pigment epithelial cell comprises the following steps of S1: culturing human pluripotent stem cells; S2: differentiating the human pluripotent stem cells obtained in S1 into retinal pigment epithelial precursor cells; S3: differentiating the retinal pigment epithelial precursor cells obtained in S2 into immature retinal pigment epithelial cells; and S4: differentiating the immature retinal pigment epithelial cells obtained in S3 to obtain retinal pigment epithelial cells. The preparation method can differentiate pluripotent stem cells rapidly, efficiently and simply to obtain retinal pigment epithelial cells. In addition, no system containing serum or a serum substitute is used in the preparation method provided by the invention. Thedifferentiation efficiency is high and the differentiation effect is stable. The purification method is simple and the obtained cell has high purity.

The invention relates to the cell transdifferentiation technology in the field of cell research, in particular to a method for inducing neuroblastoma cells to be differentiated to nerve cells. The neuroblastoma cells are cultured for 3-7 days through an inducing culture medium, and then the nerve cells are obtained; the inducing culture medium is obtained by adding retinoic acid to a neural stem cell condition culture solution to 10 micrometers; a complete culture medium including a DMEM/F12 basic cukture mediam, B27, bFGF, EGF, heparn sodium, L-glutamine and mycillin is used for culturing the neural stem cells, the complete culture medium is replaced by a half after culturing is conducted for three days, and the replaced culture solution is the neural stem cell condition culture solution. By means of the method, the differentiation efficiency is improved, and the maturity of the cells and growth of neural synapses of nerve cells are also improved. The cell apoptosis in SH-SY5Y treated through RA can be inhibited.