132results about How to "Good differentiation effect" patented technology

The invention relates to a method for separating mesenchymal stem cells from placenta. The method comprises the following steps: (a) taking placental cotyledon, and fully washing by using a phosphate buffer solution (PBS) to remove residual blood from the placenta; (b) cutting the placental cotyledon into blocks, adding a PBS containing tissue digestive enzyme, and incubating and digesting at 37DEG C; (c) filtering the tissue blocks by using a copper net, and grinding if necessary to promote filtration; (d) centrifuging the collected filtrate, separating mononuclear cells, suspending the obtained cells by using a mesenchymal stem cell (MSC) culture medium, and culturing in a 5 percent CO2 incubator at 37DEG C; and (e) after the dispersed cells form clones, selecting the clone cells, respectively culturing by using an MSC culture medium, and after the cells are fused, performing digestion and passage by using pancreatin to obtain the mesenchymal stem cells of the placenta. By the method, high purity mesenchymal stem cells of the placenta can be obtained.

The invention relates to a method for freezing and thawing placental whole cells and separating and expanding stem cells. The method comprises the steps as follows: disinfecting and washing a placenta tissue; cutting off placental lobules from the tissue, and carrying out digestive treatment for 20 min; preparing a placenta tissue freezing solution for standby application; adding whole cells obtained by digestive treatment and the freezing solution into a freezing tube, refrigerating for 0.5 h at the temperature of 4 DEG C, freezing for 1 day at the temperature of subzero 80 DEG C, and freezing in liquid nitrogen for standby application; taking out the placental whole cells from the liquid nitrogen when needed, thawing in a thermostatic water bath, carrying out drop-method washing by a culture medium for mesenchymal stem cells, removing red cells by a red cell lysis solution, and expanding the mesenchymal stem cells by the thawed placental whole cells through cell culture and cell passage. According to the method, the frozen placenta tissue can be effectively protected and is convenient to thaw and use; and the method is in particular suitable for separating and expanding the mesenchymal stem cells after thawing the frozen placental tissue.

The invention relates to a method for separating and amplifying a mesenchymal stem cell from a fresh tissue of an umbilical cord. The method comprises the following steps of: sterilizing and cleaning an umbilical cord tissue; shearing the tissue into a block shape; carrying out digestion treatment for 1-2.5 hours and ending the digestion; cleaning the cell obtained by digesting to obtain a cell suspension; adding the cell suspension into a T25 cell culture bottle for culturing; supplementing after culturing the cell suspension for 3-6 days and continuously culturing; carrying out full liquid change for the first time when the cell suspension is cultured for 8-10 days and then carrying out one full medium change every one to three days; when the fusion rate of anchorage-dependent cells in a plate reaches about 50-70 percent, enabling the anchorage-dependent cells to be separated from the bottom of the T25 cell culture bottle by using digestive enzyme; removing supernate by centrifuging, adding a mesenchymal stem cell culture medium for re-suspending the cell, inoculating the cell to the T25 cell culture bottle for carrying out passage and amplifying culture; and then changing liquid once every 1-3 days until the fusion rate reaches 70-90 percent and obtaining the umbilical cord mesenchymal stem cell. The method disclosed by the invention can be effectively used for separating and augmenting the mesenchymal stem cell.

The present invention relates to an application of iridoid in the preparation of anti-osteoporosis medicines, which belongs to the medicines technical field. The invention provides an application of iridoid which is shown in a general formula I in the preparation of medicines used for treating and/or preventing osteoporosis. Experiments found that ALP activity in UMR106 cells is taken as guidance for tracing and separating an extract of iridoid possessing general formula I, the extract and each separated component are researched from the aspects of osteoblast propagation and antioxidation activity, the iridoid possessing the general formula I has the effects of substantially promoting alkaline phosphatase activity and osteoblast differentiation, so that the iridoid has anti-osteoporosis effect and can be used for preparing the medicines used for treating or preventing osteoporosis. The component of specnuezhenide and a compound G13 possesses the effects of substantially promoting the alkaline phosphatase activity and osteoblast differentiation, and the specnuezhenide has effects for obviously promoting UMR-106 cell proliferation, a better effect is obtained when the specnuezhenide is used for anti-osteoporosis effect.

The invention relates to a method for preparing a chitosan-silk fibroin composite nano-fiber multifunctional patch for promoting myocardial tissue regeneration and monitoring stem cells. The method comprises the following steps: preparing a cellulose nano-fiber basal plate by an electrostatic spinning technology; alternately assembling positively charged chitosan (CS) and negatively charged silk fibroin (SF) to the surface of nano-fibers layer by layer by adopting a layer-by-layer assembly technique, and assembling 5.5-10.5 layers to form a CS-SF composite nano-fiber membrane; and planting seed cells of adipose tissue-derived stromal cells or cardiac progenitor cells labeled by green fluorescent protein and firefly luciferase on the surface of the CS-SF composite nano-fiber membrane, preparing the patch by three-dimensional co-culture. The patch has excellent biocompatibility, can be used as a cell vector, has an effect of resisting oxidative stress to improve the survival rate and treatment efficiency of stem cells, and is capable of evaluating the number, distribution and function states of transplanted stem cells, effectively preventing occurrence of post-myocardial infarction heart failure and reducing the death rate of ischemic cardiomyopathy.

The invention relates to a method for separating mesenchymal stem cells from placentas and a digestive enzyme composition adopted in the method. The method includes: (a) completely flushing placental lobules with PBS buffer solution to remove residual blood in the placentas; (b) cutting the placental lobules into blocks, adding into tissue digestive enzyme containing PBS buffer solution, and performing incubation digestion at 37 DEG C; (c) filtering the tissue blocks through a copper screen, and grinding to promote filtration in necessity; (d) centrifuging collected filtrate, separating mononuclear cells, adopting an MSC culture medium for suspension of the obtained cells, and culturing in a 5% CO2 incubator at 37 DEG C; (e) after cloning of dispersed cells is formed, picking each clone cell, respectively culturing with the MSC culture medium, and after cell fusion, performing trypsinization passage to obtain the mesenchymal stem cells. By adoption of the method, high-purity mesenchymal stem cells can be obtained, andn low cost and high cell yield are realized.

The invention relates to a method for separating and amplifying mesenchymal stem cells from an umbilical cord. The method comprises the following steps of: disinfecting and cleaning an umbilical cord tissue; cutting the tissue into pieces, and paving in another cell culture plate to ensure that the tissue pieces are air-dried until the tissue is attached to the plate; culturing the umbilical cord tissue in a culture medium special for the mesenchymal stem cells, supplementing/replacing liquid and clearing all umbilical cord tissue pieces when the umbilical cord tissue is cultured for 11 to 13 days, and continuing to culture; completely replacing liquid every 1 to 3 days later on; separating wall attaching cells from the bottom of the plate by using digestive enzyme when the fusion rate of the wall attaching cells in the plate reaches about 50-70 percent; centrifuging and removing a supernatant, adding into the culture medium special for the mesenchymal stem cells, suspending the cells again, and inoculating in a T25 cell culture bottle for subculture and amplification culture; and replacing liquid once every 1 to 3 days later on until the fusion rate reaches 70-90 percent to obtain the mesenchymal stem cells. The method can be effectively used for separating and amplifying the mesenchymal stem cells.

The invention relates to compound emulsifiable paste for treating acne and a preparation method thereof, in particular to a medicament for treating acne and a preparation method thereof. The compound emulsifiable paste solves the problems of long treatment course and skin oxidation, dim skin and formation of color spots in the healing process in the conventional medicament for treating the acne. The compound emulsifiable paste is prepared from clindamycin hydrochloride, metronidazole, vitamin A, vitamin C, an emulsifying agent, stearic acid, albolene, glycerol monostearate, lanolinum, glycerol and purifying water. The preparation method comprises the following steps of: adding the vitamin A into uniform solution prepared from the stearic acid, the albolene, the glycerol monostearate and the lanolinum to obtain an oil phase; dissolving the glycerol and the emulsifying agent into the purifying water, adding the clindamycin hydrochloride, the metronidazole and the vitamin C, and stirring uniformly to obtain a aqueous phase; and adding the oil phase into the aqueous phase, stirring, homogenizing and standing the mixture to obtain the compound emulsifiable paste for treating the acne. The treatment period of the compound emulsifiable paste is between 14 and 24 days, skin is bright and clean after healing, and the compound emulsifiable paste can treat the acne, seborrheic dermatitis, acne rosacea and folliculitis.

The invention relates to a method for preparing CD34 positive cells from umbilical cord mesenchymal stem cells. The method specifically comprises the following steps: (a) providing mesenchymal stem cells from an umbilical cord; (b) performing primary culture on the mesenchymal stem cells; (c) performing subculture on the mesenchymal stem cells; (d) in a subculture process, adding an inductive agent in a culture medium for inducing the mesenchymal stem cells; (e) obtaining the CD34 positive cells. The invention also relates to cell products of the CD34 positive cells, prepared through the method and application of the cell products. The method disclosed by the invention has excellent technical effects described in the description.

The invention discloses a fast breeding method for dendrobium candidum protocorm tissue culture. The method comprises the following steps that 1, explants are cleaned and sterilized; then, the explants are inoculated into an induction medium to perform induction culture; 2, the protocorm inducted in the first step is inoculated into a liquid proliferation culture medium to perform suspension shaking culture; 3, the protocorm obtained in the second step is transferred into a differentiating culture medium to be cultured for obtaining differentiated seedlings; 4, the seedlings with the height being 1 to 2cm in the third step are transferred into a rooting culture medium for rooting. The method has the advantages that the proliferation times can be improved; the industrial fast breeding can be realized. The dendrobium candidum protocorm cultured by the method has high differentiation rate; the dry weight and fresh weight content is improved; the transplanting survival rate is correspondingly improved.

The invention provides mesenchymal stem cell injection liquid and a preparation method, which aim to solve the problem of how to use a mesenchymal stem cell to efficiently repair and improve damaged skin and fundamentally delay and stop skin cell aging. The injection liquid provided by the invention is prepared from the mesenchymal stem cell, DMSO, EGF, glutathione, vitamin C, human serum albumin,a ginkgo biloba extract and a menstruum. The mesenchymal stem cell is obtained through carrying out hungry culture on P2 to P5-generation mesenchymal stem cells and stimulating through an epidermal growth factor. The obtained mesenchymal stem cell has a good differentiative capacity and a good proliferation capacity, has a better differentiation potential, and is synergized with the DMSO, the EGF, the glutathione, the vitamin C, the human serum albumin and the ginkgo biloba extract, so that skin wrinkles and hyperpigmentation can be remarkably reduced.

The invention relates to a method for treating premature ovarian failure by virtue of placenta mesenchymal stem cells and a cell preparation and particularly relates to the cell preparation on one hand. The cell preparation is cell suspension prepared by mixing mesenchymal stem cells such as placenta mesenchymal stem cells into a 0.9% sodium chloride solution and is particularly prepared by the steps of transferring mesenchymal stem cells obtained through cell passage into a centrifuge tube, carrying out centrifuging to remove supernatant, adding the 0.9% sodium chloride solution, and carryingout resuspension, so as to obtain the cell preparation. The mesenchymal stem cells are prepared by the steps of processing placental lobules, carrying out digestion and termination on mixed enzyme, collecting primary cells, carrying out cryopreservation on the primary cells, carrying out thawing, passage, detection and cryopreservation on the cells, and relating with a database. The cell preparation prepared by virtue of the method presents excellent biological effect in the treatment of the premature ovarian failure.

The invention relates to canine umbilical cord mesenchymal stem cells as well as a preparation method and a cryopreservation method thereof. Specifically, the method comprises the following steps: disinfection and washing, digestion treatment, cell culture, cell passage, and cell cryopreservation. The method for preparing mesenchymal stem cells from a canine umbilical cord exhibits an excellent technical effect. The mesenchymal stem cells is beneficial for treatment of canine arthritis, fractures, muscle damage, ligament injury, cartilage damage, joint damage, cognitive dysfunction, immune-mediated diseases, dry eye, recurrence uveitis, liver disease, heart disease, kidney disease, diabetes, gastrointestinal disease, thyroid disease and skin disease.

The invention relates to a placenta mesenchymal stem cell preparation and application of the placenta mesenchymal stem cell preparation in treating sclerosis. Specifically, the invention relates to the cell preparation. The cell preparation is a cell suspension prepared by mixing and suspending mesenchymal stem cells such as placenta mesenchymal stem cells in a 0.9% sodium chloride solution and isspecifically prepared through a method which comprises the following steps of transferring the mesenchymal stem cells obtained through cell passage to a centrifuge tube, carrying out centrifuging, discarding a supernatant, and adding the 0.9% sodium chloride solution for re-suspending so as to prepare the cell preparation. The invention further relates to the application of the cell preparation in preparation of drugs for treating and/or preventing the systemic sclerosis. The invention further relates to a method for preparing the cell preparation and the placenta mesenchymal stem cells usedby the cell preparation and mixed enzyme digestive juice used in the method. The cell preparation has an excellent biological effect in the aspect of treating the sclerosis.

The invention relates to a placental mesenchymal stem cell preparation used for treating premature ovarian failure. Particularly, one the one hand, the method that mesenchymal stem cells are separatedfrom placenta tissue and cultured into the mesenchymal stem cells is provided; on the other hand, the cell preparation is provided, wherein the cell preparation is a cell suspension which is preparedin the step that the mesenchymal stem cells, such as placental mesenchymal stem cells, are mixed and suspended into a 0.9% sodium chloride solution. The cell preparation can be used for treating thepremature ovarian failure; particularly, the cell preparation is the placental mesenchymal stem cell preparation. The cell preparation is prepared by using the method including the steps that the mesenchymal stem cells obtained through cell passage are transferred into a centrifuge tube and centrifuged, liquid supernatant is removed, a 0.9% sodium chloride solution is added for resuspension, and the cell preparation is prepared. The cell preparation has an excellent biological effect on treatment of the premature ovarian failure.

The invention provides an application of a method for the autocrine secretion of an extracellular matrix by stem cells and the induction of the stem cells to become hepatocytes. The extracellular matrix is secreted by inducing bone marrow mesenchymal stem cells and removes the matrix of the bone marrow mesenchymal stem cells. The invention concretely provides the method for inducing the bone marrow mesenchymal stem cells into hepatocytes by the extracellular matrix according to actual demands. The method for generating the hepatocytes through induction has the advantages of high efficiency, practicality, high induction efficiency and good differentiation effect, and the glycogen synthesis capability and the urea synthesis capability of the hepatocytes obtained after the induction are strong.

The present invention relates to a method for isolating mesenchymal stem cells from placental vessels and used digestive enzyme compositions. The method comprises the following steps: the placenta issterilized with alcohol; Dissecting placental blood vessels from the placenta; Cut into pieces, wash with PBS, filter the residual blood stains to obtain placental vascular tissue; Adding mixed enzymesolution to digest; End digestion, filter and collect tissue fluid; The placental mesenchymal stem cells (P0) were obtained by centrifugation and cultured with DMEM-F12 basal medium was resuspended and the number and viability of nucleated cells were counted. The obtained cells are cryopreserved with cryopreservation protection solution so as to be re-cultured before use, or the passages and/or the mesenchymal stem cells are continued to be subjected to cell identification and/or detection, cryopreservation, library establishment and the like. The method of the invention can effectively improve the efficiency of isolating mesenchymal stem cells from the blood vessels of the placenta.

The invention relates to a method for preparing mesenchymal stem cells from the umbilical cord of a dog. Specifically, the method comprises the following steps: sterilization and cleaning, digestion treatment, cell culture, cell passage and cell cryopreservation. The method for preparing mesenchymal stem cells from the umbilical cord of the dog, which is provided by the invention, has excellent technical effects; the obtained mesenchymal stem cells are beneficial to treatment on arthritis, bone fracture, muscle injury, ligamentous injury, cartilage injury, joint injury, cognition impairment, immune-mediated diseases, xerophthalmia, recurrent uveitis, hepatic diseases, heart diseases, kidney diseases, diabetes, gastrointestinal diseases, thyroid diseases and skin diseases of canidae.

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 compound bone cement, a preparation method and application of the compound bone cement, and a bone repair material. The compound bone cement is prepared by mixing the followingraw materials in percentage by mass: 50-80% of solid phase powder and 20-50% of setting liquid, wherein the solid phase powder comprises phosphate and/or acidic salt of phosphoric acid; the setting liquid comprises chitosan, hyaluronic acid, organic acids and water. The compound bone cement has excellent syringeability, biological activity and bone conduction and repair ability, the set bond repair material has an excellent mechanical property, the surface of the material is easy for cell adhesion, and the cell differentiation ability cannot be influenced. The compound bone cement disclosed by the invention can be used for bone defect filling and repairing and serves as a drug carrier to be used for treating orthopedic related diseases.

The invention provides a culture method for inducing mesenchymal stem cell to differentiate for forming osteocyte under the condition without CO2. The culture method comprises the following steps of: culturing the third-generation human mesenchymal stem cell under the conditions of 5 percent of CO2 and 37 DEG C and standing overnight; after the cell is completely attached to the wall and grows until 80 percents of cell is fused, replacing an inductive culture medium and culturing under the conditions of 37 DEG C without CO2 for 14-21 days to obtain osteoblast, wherein the inductive culture medium is formed by adding fetal calf serum, penicillin, streptomycin, hexadecadrol, beta-sodium glycerophosphate and vitamin C in a basal medium and is prepared from the following components calculatedby final concentration: 0.41g/L of sodium bicarbonate, 1.5g/L of disodium hydrogen phosphate dodecahydrate, 0.07g/L of monopotassium phosphate and 8.17g/L of sodium chloride; and solvent of the inductive culture medium is an L-DMEM cell culture medium free of sodium bicarbonate. According to the novel culture medium formula provided by the invention, the mesenchymal stem cell osteoblast can be normally induced to differentiate; the differentiation effect is favorable, so that the consumption of equipment is reduced; and the novel culture medium formula is suitable for cell culture and research on induced differentiation under the condition without CO2.

The invention relates to a method for treating scleroses using a placental mesenchymal stem cell preparation. Particularly, on one hand, the invention relates to the use of the cell preparation for the manufacture of a medicament for the treatment and/or prevention of systemic sclerosis; the cell preparation is a cell suspension prepared by suspending mesenchymal stem cells such as placental mesenchymal stem cells in a 0.9% sodium chloride solution, and the cell preparation has a cell concentration of 1-10*10<6> cells/ml. On the other hand, the invention also relates to a cell preparation forthe treatment and/or prevention of systemic sclerosis, and a method for preparing the cell preparation, the method for preparing mesenchymal stem cells and the mesenchymal stem cells are involved in the cell preparation, the mixed enzyme digest is used in these methods. The cell preparation has excellent biological effect in the aspect of treating scleroderma.