842 results about "Bone marrow mesenchymal stem cells" patented technology

Disclosed are compositions and methods for augmenting bone formation by administering isolated human mesenchymal stem cells (hMSCs) with a ceramic material or matrix or by administering hMSCs; fresh, whole marrow; or combinations thereof in a resorbable biopolymer which supports their differentiation into the osteogenic lineage. Contemplated is the delivery of (i) isolated, culture-expanded, human mesenchymal stem cells; (ii) freshly aspirated bone marrow; or (iii) their combination in a carrier material or matrix.

The present invention discloses the separation method of mesenchymal stem cell in human placenta. On the basis of past separation of tissue cell, the present inventor separates from placenta mesenchymal stem cell with high purity via perfusion process based on the special anatomical structure of placenta. Identification result shows that the mesenchymal stem cell separated from placenta has the biological characteristic and polydirectional differentiation capacity as the reported mesenchymal stem cell of marrow. Owing to the infantile cell component and wide source of placenta, like cord blood, the present invention will have wide clinical application foreground.

A method for differentiating mesenchymal stem cells of bone marrow into neural cells comprises culturing the mesenchymal stem cells in a medium containing epidermal growth factor(EGF), basic fibroblast growth factor(bFGF) and hepatocyte growth factor(HGF), and the neural cells produced thereby can be employed for the treatment of a neural disease.

The present invention belongs to the field of biotechnology, and discloses a serum-free culture medium with specific chemical compositions for in vitro culture and amplification of bone marrow mesenchymal stem cells. By adding insulin, transferrin, ethanolamine, sodium selenite, growth factors, adherent factors, hormone, putrescine, inorganic salt, vitamin, albumin and antioxidant into a basic culture medium, the bone marrow mesenchymal stem cells can attach to the culture medium under a serum free condition, so the in vitro culture and amplification are realized, the potential of multi-directional differentiation is maintained, and the amplified cells can be induced to be osteoblast and lipocyte in vitro. The serum-free culture medium has the advantages that the clinic level cell products for human produced by the serum free culture medium can effectively avoid the potential risk of producing cell products by serum culture medium. The drawing appended is a photo of the confluence of the bone marrow mesenchymal stem cells cultured by the serum-free culture medium.

The invention relates to a cell model obtained after targeted knockout of a rabbit BMP2 gene based on CRISPR/Cas9 and application thereof, belonging to the technical field of molecular biology and biomedicine. According to the invention corresponding oligos are synthesized for three targeting sites of the rabbit BMP2 gene on the design principles of CRISPR/Cas9 and are constructed on px458 vectors; and a CRISPR/Cas9 system is constructed directed at the three targeting sites in rabbit mesenchymal stem cells, and the CRISPR/Cas9 system can effectively knock out the rabbit BMP2 gene, is easy to operate and has high rabbit BMP2 gene knockout efficiency. The cell model disclosed in the invention can greatly promote research related to the functions and signaling pathways of the BMP2 gene.

The invention provides icariin application in inducing stem cell in vitro orientation differentiates to several single type cells. The stem cell said includes embryonic stem cell, nerve stem cell and marrow mesenchyme stem cell. The single type cell includes nerve cell, bone cell, islet cells and endothelial cell. This invention also relates to application of the single type cell in preparing medicine of stem cell transplant curing nerve degenerative diseases, and its application in preparing cell differentiation agent that used to repair recovery injured nerve tissue, and its application in high efficiency drug effect screening model rebuilding and initial screening and estimating by using the model. The new applications of the icariin is extended in this invention, the fact of icariin contains pharmacy activity clarified, so it provides material basis for Chinese traditional medicine prevention and cure effect. The clarifying of drug effect mechanism provides reference for new medicine Chinese metical modern development with self-owned intellectual property right.

The invention relates to the field of biotechnology and mainly relates to a use of an early-generation human umbilical cord mesenchymal stem cell exosome in promotion of human bone mesenchymal stem cell growth. The use method is a method for promoting bone mesenchymal stem cell growth. The invention also relates to an extraction method of a human umbilical cord mesenchymal stem cell exosome. The early-generation human umbilical cord mesenchymal stem cell exosome and human bone mesenchymal stem cells are co-cultured so that human bone mesenchymal stem cell growth is obviously promoted, cell doubling time is shortened and a ratio of an S period in a cell period is increased.

The invention discloses a preparation method of original mesenchymal stem cells. In the invention, the mesenchymal stem cells are prepared by adopting fresh human umbilical cord, extracting Huatong glue, digesting by collagenase, and separating and cultivating, and the mesenchymal stem cells are suitable for cryopreservation. The invention also relates to the original mesenchymal stem cells prepared by the method, and the original mesenchymal stem cells are verified as mesenchymal stem cells by the identification of biological phenotype according to standard formulated by the International Society of Cytotherapy. The multiplication capability and the purity of the mesenchymal stem cell are both higher than those of bone mesenchymal stem cells; and the amount of the released growth factorsand cell factors is higher than that of the bone mesenchymal stem cells.

The invention provides an inducing method and inducing culture medium for differentiation of bone marrow mesenchymal stem cells into osteoblasts in vitro. The inducing culture medium is composed of 1*10<-8> mol/L of dexamethasone, 50 mu mol/L of ascorbic acid and 10 mmol/L of sodium beta-glycerophosphate; and solvent is a supernatant of a sclerite complete culture medium and comprises 10% of fetal calf serum, 100 U/mL of penicillin, 100 mg/L of streptomycin, a mixture of DMEM culture fluid and F12 culture fluid and multiple growth factors secreted by bone cells in the sclerite culture process. According to the invention, bone marrow mesenchymal stem cells of a mouse are purified by replacing the cell culture fluid through an adherent cell passage method, the obtained cells of the first generation are induced, and the supernatant of the sclerite complete culture medium cultured for 72-96 hours is used as the solvent of osteoblast differentiation inducer, thereby obviously improving the in vitro osteogenic differentiation efficiency of bone marrow mesenchymal stem cells.

The invention provides an integrated bone-cartilage repair scaffold. The integrated bone-cartilage repair scaffold is of an integrated structure composed of a subchondral bone repair layer, a middlelayer and a cartilage repair layer, wherein the subchondral bone repair layer is fashioned porous calcium phosphate biological ceramic, the middle layer is sulfydryl-hyaluronic acid hydrogel, the cartilage repair layer is composed of I-type collagen hydrogel and chondrocytes or mesenchymal stem cells, and the middle layer is located between the subchondral bone repair layer and the cartilage repair layer and in a porous structure of the fashioned porous calcium phosphate biological ceramic to separate the subchondral bone repair layer from the cartilage repair layer. The invention further provides a preparation method of the integrated bone-cartilage repair scaffold. The repair scaffold can prevent blood vessels from invading a cartilage layer, so that the tightness and the stability of connection between a bone layer and the cartilage layer are improved, biological bonding of the bone layer and the cartilage layer is achieved, and thus a good repair effect is achieved.

The invention discloses a tissue-engineered bone. The tissue-engineered bone is multilayer cell sheet lamination compound with a three-dimensional capillary network, the multilayer cell sheet lamination compound consists of n laminated bone marrow mesenchymal stem cell sheets and vascular endothelial cells among all bone marrow mesenchymal stem cell sheet layers, and n is in a range of 3-8. A preparation method of the tissue-engineered bone comprises steps as follows: firstly, a photosensitive semiconductor structural layer is prepared on the surface of a cell culture dish, so that a photosensitive cell culture dish is obtained; then bone marrow mesenchymal stem cells are cultured with the photosensitive cell culture dish, and the bone marrow mesenchymal stem cell sheets are obtained; finally, the multilayer cell sheet lamination compound consisting of multiple layers of the bone marrow mesenchymal stem cell sheets and the vascular endothelial cells is constructed and cultured to obtain the tissue-engineered bone. The tissue-engineered bone purely consists of homologous cells, so that pollution caused by immunological rejection and stent degradation is reduced, and the tissue-engineered bone is significant in bone defect repair and early vascularization; the method is simple, easy to implement and convenient to popularize.

The invention discloses a cartilage tissue engineering repair bracket and a preparation method thereof, and belongs to the cartilage tissue repair field. The bracket comprises natural high polymer water-gel and decalcified bone matrix which is coupled with affinitive peptides of mesenchymal stem cells on surface; the affinitive peptides of the mesenchymal stem cells are coupled on the surface of the decalcified bone matrix to obtain the cartilage tissue engineering repair bracket with special mesenchymal stem cell affinity. By being placed in a cartilage defective region, the cartilage tissue engineering repair bracket can collect a plurality of BMSC (bone marrow stromal cells); and in the bracket, under mating effect of the natural high polymer water-gel and the decalcified bone matrix, the BMSC can be retained in the cartilage defective region for a long time, so that the cartilage repair has a longer curative effect period. The invention further discloses a preparation method of the bracket, wherein the natural high polymer water-gel aqueous liquor is injected in the decalcified bone matrix which is coupled with affinitive peptides of mesenchymal stem cells on surface to carry out gelation reaction to obtain the bracket with higher mechanical strength and inducibility. The method is simple and high in practicability.

The invention relates to a medical dressing with bioactivity and a preparation method of the medical dressing and belongs to the technical field of biomedical engineering and material science. The medical dressing with the bioactivity is used for covering skin defect wounds caused by burning, scalding, operation, wounds and diseases and inducing wound repair and is prepared as follows: one or more of embryonic stem cells, umbilical cord blood stem cells, amniotic fluid stem cells, peripheral blood stem cells and bone mesenchymal stem cells of humans or animals are cultured in vitro, stem cell growth factors secreted in the logarithmic growth phase are collected and mixed in proportion with one or more of I type collagen, III type collagen, chitosan, hyaluronic acid, chondroitin sulfate and sodium alginate, thin-layer sponge is taken as an inner layer, a porous high-polymer material film outer layer prepared from a mixture formed by one or more of PLA (polylactic acid), PGA (polyglycolic acid), PLGA (poly(lactic-co-glycolic acid)) and PCL (polycaprolactone) is attached, the product is frozen-dried and cut, and the medical dressing with the bioactivity is formed and used for treating skin defect wounds caused by burning, scalding, operation, wounds and diseases and inducing wound repairing.

The present invention discloses a stem cell culture medium and its applications as well as a stem cell culture method. The said stem cell culture medium contains no serum. The said stem cell culture medium contains amino acids, vitamins, salts, lipids, cytokines and protein polypeptides. The said stem cell culture medium is suitable for rapid culture of stem cells derived from human and mammalian tissues, including but not limited to, adipose mesenchymal stem cells, bone marrow mesenchymal stem cells and umbilical cord blood stem cells. The said culture medium can increase the proliferation speed of the stem cells 3-5 times, without any affects on their differentiation potentials. Comparing the said stem cell culture medium to a routine culture medium, the said culture medium is not only able to proliferate stem cells derived from different sources more rapidly and achieve more proliferation generations, but also keep their differentiation potentials well.

The invention discloses a tissue engineered bone compound which comprises porous perforated between aperture artificial bone bracket and mesenchymal stem cells implanted in hole and transfected with vascular endothelial growth factor gene. The invention also discloses the application of the tissue engineered bone compound as material for bone defect repair. The inventive tissue engineered bone compound can make seed cell exert osteogenesis, express and secrete VEGF, thereby promoting vasal formation and growth, providing material basis for new bone formation, in favor of neogenesis of vascularized tissue engineered bone tissue, improving effect for repairing bone defect.

The present inventors revealed the following for the first time in the world:

• 1) a large amount of bone marrow-derived cells are mobilized to grafted skin;
• 2) the mobilized bone marrow-derived cells are differentiated into any of dermal fibroblasts, adipocytes, muscle cells, vascular endothelial cells, and epidermal keratinocytes in the grafted skin, and the mobilized bone marrow derived cells include bone marrow-derived mesenchymal stem cells;
• 3) the factors which mobilize bone marrow-derived mesenchymal stem cells from peripheral blood to the grafted skin are HMGB1, HMGB2, and HMGB3 released from the necrosed tissue of recipient skin;
• 4) purified HMGB1, HMGB2, and HMGB3 promote the migration of mesenchymal stem cells isolated and cultured from bone marrow;
• 5) activators containing HMGB1 which allows the migration of bone marrow mesenchymal stem cells can be conveniently purified from several organ extracts including skin, brain, and heart;
• 6) activators which allow the migration of bone marrow mesenchymal stem cells can be conveniently extracted from cultured cells; and
• 7) a heparin-column purified fraction of skin extract mobilizes a large amount of bone marrow-derived cells in case of brain injury.

The invention provides a kit and method for inducing differentiation from bone mesenchymal stem cells to osteoblasts. The kit comprises an osteoblast inducing culture solution and an osteoblast identification solution, wherein the inducing culture solution comprises a cell culture solution A, a cell culture solution B, a cell culture solution C, a cell culture solution D and a cell culture solution E; the osteoblast identification solution comprises a cell immobilizing solution, coloring agents and a detergent; the cell culture solution A is an alpha-minimum essential medium (MEM) liquid culture medium; the cell culture solution B is fetal bovine serum; the cell culture solution C is a dexamethasone solution; the cell culture solution D is a beta-sodium glycerophosphate solution; the cell culture solution E is a vitamin C solution; the cell immobilizing solution is 4% paraformaldehyde; the coloring agents include sodium thiosulfate, silver nitrate and neutral red; and the detergent is double distilled water.

The invention provides a preparation method of a multifunctional layered joint cartilage support. The multifunctional layered joint cartilage support capable of simulating the biological property andthe mechanical property of natural cartilage is prepared by taking hydrogel as a support material, taking mesenchymal stem cells, cartilage cells and osteoblasts as inducing and developing objects andby adopting a 3D printing technology. The multifunctional layered joint cartilage support can accurately simulate the internal tissue form and the contour of each layer of the natural cartilage and can realize that different layers have different mechanical and biological characteristics. The prepared artificial cartilage support cells are derived from patients, the support serves as a degradablehydrogel material and the rejection reaction after implantation is greatly reduced. Various nutritional substances and growth factors are mixed in the hydrogel support, so that the sustained releaseeffect can be achieved and regulation and control of the growth and development of the artificial cartilage can be realized.

The invention relates to a preparation method and application of collagen scaffold composite bone marrow-derived mesenchymal stem cells (BMSCs). The preparation method comprises the steps of preparing single cell suspension after trypsinizing the BMSCs, uniformly dropwise adding the single cell suspension to a collagen scaffold, putting the collagen scaffold into an incubator to be cultured, and adding an L-DMEM complete medium to continue culture, thus obtaining the collagen scaffold composite BMSCs. The preparation method has the advantages that the following defects are overcome: endometria are seriously injured as intrauterine adhesion is mechanically separated by adopting hysteroscopic surgery, intrauterine devices or anti-adhesion materials are put after the surgery and estrogens are given after the surgery to promote intima growth; the problem of intima scars can not be solved; functional intima repair can not be achieved; adhesion is very easy to happen again. As active ingredients for treating serious endometrium injury, the BMSCs are convenient to obtain, secrete growth factors to improve the local microenvironment and immunoregulation, have good biocompatibility, degradability and safety, promote scarred endometrium repair and increase the intima thickness and local blood vessel density.

The invention discloses an application of umbilical cord mesenchymal stem cells (MSCs) in the acute myocardial infarction cell transplantation therapy; the application is suitable for ST-segment elevation acute myocardial infarction within one mouth or non ST-segment elevation acute myocardial infarction cases, a vascular remodeling is carried out to a patient who is attacked by the disease withinsix hours, after about four to six days, coronary perfusion is carried out to the patient; and the other patients who choose date to carry out vascular remodeling operation, the coronary perfusion iscarried out at the same time. The treating effect in the invention is superior to the bone mesenchymal stem cells transplantation.

The invention relates to a cell injection for treating bone injury and a preparation method thereof, belonging to the field of tissue engineering. The injection is a compound formed by mesenchymal stem cells from autologous bone marrow, temperature sensitive gel and effective trophic factors separated and extracted from autoblood. The preparation method comprises the steps of: after extracting the bone marrow of a patient, separating and extracting the mesenchymal stem cells from the bone marrow, adding no animal serum in the amplification culture process, and mixing 1*(105-108) cells with 1-5ml of biological scaffold material to form the injectable compound. The cell injection for treating bone injury is mainly used for treating bone injuries including osteoarthritis, femoral head necrosis and delayed union and disunion of bone fracture, is convenient to use, has notable effect without immunological rejection, and can alleviate the great pain of patients caused by bone injury.

The invention provides a method for inducing mesenchymal stem cell in the bone marrow to obtain the nerve prosoma cell, which comprises the following steps: doing generation culture for the separated and purified mesenchymal stem cell in the bone marrow, switching stable generation mesenchymal stem cell into induction culture medium, inducing and culturing for 7-20 d, obtaining nerve prosoma cell. The induction culture medium is basic culture medium which is charged with growth factor, wherein the growth factor mainly comprises an epiderm growth factor (EGF), a base fibroblast growth factor (bFGF), an insulin growth factor-1 (IGF-1) and a nerve nourishment factor 3 (NT-3). The invention provides a method for inducing mesenchymal stem cell in the bone marrow to obtain the nerve prosoma cell which can be differentiated into nerve cell and the nerve prosoma cell can be switched into inner ear hair cell and inner ear supporting cells, which provides experiment evidence for cochlear cell transplantation therapeutic sound perception nerve deafness and solves the problem of the transplantation cell resource.

The invention provides traditional Chinese medicinal exosome-loaded compound dressing with effects of promoting wound healing and regeneration and a preparation method of the compound dressing. The preparation method comprises the following steps: preparing methacrylate esterified gelatin by a reaction of gelatin and methacrylic anhydride; preparing a traditional Chinese medicinal exosome throughdifferential centrifugation; combining the prepared exosome with the methacrylate esterified gelatin, adding SDF-1 and photosensitizer into the mixture to prepare a pre-gel system; and forming the compound dressing through ultraviolet crosslinking. The dressing is novel photosensitive dressing of a compound traditional Chinese medicinal exosome, the compound can be formed through ultraviolet crosslinking in vitro or in-situ crosslinking on a wound after the pre-gel system is formed. The compound dressing is quick in crosslinking, is simple and convenient, can be used for effectively protectinga wound area, inducing migration of mesenchymal stem cells and promoting wound heating and regeneration with the traditional Chinese medicinal exosome, and can effectively solve the problem in current wound healing and regenerating process. Therefore, the compound dressing has a wide application value in wound healing and regeneration.

A method for preparing chondroblast of bone marrow mesogalia stem cell with three dimensional cultivation and induction includes steps of placing bone marrow mesogalia stem cell into suspending culture device preset with culture medium and microcarrier for suspending cultivation; cloning abovesaid stem cells with agitation, adding induction culture medium of chondroblast in culture meidum containing harvested cytodex3 microcrrier and abovesaid stem cells gromn on it; and carrying out induction for chondroblast assembly from abovesaid stem cells.