218results about How to "Strong differentiation ability" patented technology

The invention discloses a preparation method and application of an antibacterial bio-ceramic film with the titanium or titanium alloy surface containing copper and belongs to the technical field of treating metal surfaces. The preparation method comprises the following steps: preparing a Ca, P and Cu containing porous hard ceramic layer under a single square wave pulsed power supply mode; and carrying out hydrothermal treatment or alkali treatment to obtain the CuO/HA antibacterial bio-ceramic film. According to the preparation method disclosed by the invention, the prepared ceramic layer and a matrix are in metallurgical bonding and are good in bioactivity and obvious in antibacterial effect; the prepared antibacterial bio-ceramic film is suitable for dental implant in the field of oral cavities as well as bone union plates, bone nails and the like of orthopaedics; and the ceramic layer is good in protein adsorbability and adhesion, increment and differentiation abilities of cells and is capable of greatly shortening the bonding time of the traditional medical implanting material and bones.

The invention discloses a stem cell medium and an application thereof. The medium comprises a basic medium, fetal bovine serum, cytokine or protein polypeptide, vitamin and lipid. The medium provided by the invention can quickly amplify stem cells and does not influence the potential of the stem cells, and the amplification velocity of the stem cells is improved by 3-5 times than a conventional medium; and moreover, the medium can be used for culturing the stem cells of various tissues and has excellent applicability, the cultured stem cells have strong differentiation capability and can be differentiated into cells with various functions; therefore, the stem cell medium has high scientific and research and medical application values.

The invention relates to a reprogrammed culture medium and a culture method of a reprogrammed induced multipotential stem cell. The reprogrammed culture medium is composed of a DMEM-F12 basal culturemedium and additive components, wherein the additive components comprise 60-180 microgram/mL of L-ascorbic acid, 5.3-74 micromole/L of hydrocortisone, 3-89 nanogram/mL of sodium selenite, 8-23 micromole/L of Optiferrin, 0.5-7.4 micromole/L of retinyl acetate, 40-60 nanogram/mL plant-derived recombinant human alkaline growth factors, 8-12 microgram/mL of IGF, 0.2-0.6 microgram/mL A-83, 2-6 micromole/L of CHIR99021 and 100-450 micromole/L of sodium butyrate. The use of the reprogrammed culture medium not only can improve the safety of clinical use, but also can improve the efficiency of adult cell reprogramming to induce the multipotential stem cell.

Disclosed herein are methods and compositions for replenishing injured and/or damaged cardiomyocytes in a subject by inducing, increasing, and/or enhancing the differentiation of endogenous stem and progenitor cells in the subject.

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 discloses a culture method for restoring totipotency of a mesenchymal stem cell. The method for restoring totipotency of a mesenchymal stem cell, provided by the invention, comprises the following steps: 1) performing hanging-drop culture on an in-vitro mesenchymal stem cell with reduced totipotency, thus obtaining a hanging-drop cultured cell; and 2) further performing suspension culture on the hanging-drop cultured cell, thus realizing totipotency restoration of the mesenchymal stem cell. The experiment of the invention proves that, according to the method for restoring totipotency of a mesenchymal stem cell (specifically 3D culture), after the mesenchymal stem cell (in-vitro mesenchymal stem cell with reduced totipotency) subcultured multiple times is subjected to 3D culture treatment, the increased volume is gradually restored, the cell viability is enhanced, the cloning capability and differentiation capability are enhanced, and the paracrine action is also improved, thereby activating the aged mesenchymal stem cell and restoring the characteristics of the stem cell.

The invention provides a tissue engineering meniscus scaffold and a preparation method thereof. The preparation method comprises the following steps: placing a polycaprolactone material into a nozzle of a fused deposition modeling three-dimensional printer to be heated to 120-140 DEG C, and preparing for printing at the air pressure of 600-1000 kPa; setting the printing speed of the fused deposition modeling three-dimensional printer to be 0.6-0.75 mm/s, the fiber diameter to be 300-320 [mu]m and the fiber interval to be 200-300 [mu]m and printing a tissue engineering meniscus scaffold; using cobalt-60 for irradiation and sterilization treatment. As the fused deposition modeling method is adopted to print the tissue engineering meniscus scaffold with an appropriate pore structure, an appropriate fiber diameter and appropriate biomechanical strength, the survival rate and the reproduction rate of inoculated cells and autologous in-growth cells of implanted regions are favorably increased, the fibrous cartilage differentiation rate can be increased, and newly generated meniscus tissues have favorable functions.

The invention discloses an umbilical cord mesenchymal stem cell preparation and a preparation method and application thereof. Through exploration and verification by a large quantity of experiments, the 3rd-10th-generation umbilical cord mesenchymal stem cells are selected as raw materials, and by a method of dispersing tissue by a mechanical method and then performing treatment with mixed enzymes containing collagenase I, collagenase II and trypsin for a short time for many times, the vitality of the separated umbilical cord mesenchymal stem cells is guaranteed. A culture medium special for the umbilical cord mesenchymal stem cell preparation, containing DMEM/F12 (without phenol red), 10% FBS, 20 kinds of amino acids and 8 kinds of vitamins, is adopted, mixed liquor of repeated cell culture supernatant and cell lysate supernatant is collected, more umbilical cord mesenchymal stem cell active protein and active factors are obtained, the preparation cost is reduced, and industrial requirements for a stem cell product preparation are met. The prepared umbilical cord mesenchymal stem cell preparation can promote proliferation of epidermic cells and skin fibroblast, is safe and is free from toxic and side effects, an allergy does not exist, and when the umbilical cord mesenchymal stem cell preparation is applied to feature-beautifying skin-care products, the effects are notable.

The invention provides a method for rapidly separating and extracting hUC-MSC (human Umbilical Cord mesenchymal stem cells). The method comprises the following steps: taking the freshly collected umbilical cord tissue of a healthy newborn baby, carrying out on-ice transportation on the freshly collected umbilical cord tissue in umbilical cord storage transportation liquid containing double antibodies, carrying out cleaning and disinfection by adopting 75% alcohol and normal saline, removing blood vessels, carrying out blunt dissection on wharton jelly, carrying out mechanical pulverization, treating the obtained product I by adopting red blood cell lysis buffer for 3 min, digesting the obtained product II by adopting IV collagenase, screening the obtained product III by adopting a 100-200-mesh sieve, carrying out suspension culture on the obtained product IV by adopting a serum-free medium, wherein the liquid is changed every 3-5 days, taking supernatant, detecting cell pollution, after the adherent rate in a plate reaches 30-70%, carrying out trypsinization, carrying out centrifugation, collecting cells, carrying out passage amplification, carrying out merging when the cell merging rate reaches 90% or above, collecting the cells, carrying out cryopreservation on the cells, and detecting the biological characteristics of hUC-MSC.

The invention provides a stem cell injection for treating osteoarthritis cartilage defect and preparation and application thereof. The injection comprises fifth-generation umbilical cord mesenchymal stem cells and a liquid solvent, wherein the fifth generation umbilical cord mesenchymal stem cells are suspended in the liquid solvent, and the concentration is (2.0-3.0)x10<7> cells/ 4.5-5.0ml of the liquid solvent. The liquid solvent is normal saline, or autologous platelet-rich plasma, or a mixed solution of normal saline and human albumin, or a mixed solution of normal saline and autologous platelet-rich plasma. The human albumin accounts for 5-8% in volume by percentage. The normal saline accounts for 10% or below in volume by percentage. The injection is administered by intravenous injection or bone joint cavity local injection or the combination of the two, the cell injection amount of the intravenous injection is (1.0-1.5)x10<8> cells/ one time, and the cell injection amount of the bone joint cavity injection is (2.0-3.0)x10<7> cells/ one time/ one knee. The effect of the stem cell injection for treating the integrity degree of osteoarthritis cartilage is obviously superior to that of a traditional hyaluronic acid (HA) method.

The various embodiments herein provide a method of co-culturing human endometrial stem cells and rat embryonic tooth bud cells to obtain ameloblast cells. The human endometrial stem cells are isolated from human females between the ages of 18-40 and cultured. The hEnSCs are identified using flowcytometry. The rat embryonic tooth bud cells are isolated from rat embryos and cultured. After the hEnSCs and rat embryonic tooth bud cells reach a desired level of cell growth, the hEnSCs are transferred to culture plate inserts and the rat embryonic tooth bud cells are transferred to a culture plate. The hEnSCs and rat embryonic tooth bud cells are co-cultured to obtain ameloblast cells. Immunocytochemical analysis is used to characterize the differentiated ameloblast cells. Quantitative real time PCR (qRT-PCR) is used to detect the presence of Ameloblastin, Amelogenin, Amelotin and Cytokeratin 14 proteins in differentiated ameloblast cells.

The invention discloses a Cyclocarya paliurus cutting seedling growing method comprising the following steps: 1, cuttingwood selecting; 2, cuttingwood processing; 3, cuttingwood planting; 4, post-planting management; the method uses specially prepared matrix, and the matrix has excellent physicochemical performance so as to provide good environment conditions for cuttingwood growth; nitration soup immersion and electric field alternative repeatedly treatment are carried out for the cuttingwood root, thus enhancing root cell splitting and differentiation capability, and improving growth vitality and anti adversity performance of the cuttingwood; combining with matrix effect and post-planting management, the novel method can largely improve cuttingwood survival rate, and the seedling is tidy and prosperous in growth vigor, thus reducing incubation time for 7-10 days when compared with same specification cutting seedlings; the method is suitable for large-area popularization plantation, and very strong in market competitiveness.

Enhanced postnatal adherent cells and a preparation method therefor are provided. The preparation method of enhanced postnatal adherent cells can increase the yield and the proliferation rate of adherent cells from placental tissues; and prepare adherent cells, which secrete proteins effective for neurological diseases and have an improved ability for movement to damaged tissues.

The invention discloses a method for increasing the grafting survival rate of a peach tree. The method is characterized by comprising the following steps of 1, branch selection, wherein the 4-5-years-old peach tree is adopted as a parent tree, and the lower portions of soft branches are bilaterally cut by 30 degrees; 2, cion treatment, wherein a cion branch is soaked in a sterilization solution and a growth promoting solution, and then soaked in plant extract; 3, stock treatment, wherein a stock is firstly treated through carbendazim solution and growth promoting solution spraying, and the wound portion of the stock is sealed with plant glue liquid; 4, grafting, wherein the stock with a wrapping layer removed and the cion are combined, a growth promoting solution and plant extract are sprayed to the combination portion, then plant glue liquid is used for repeating coating, and the combination portion is wrapped by gauze and a preservative film; 5, daily management, wherein during woundhealing, the peach tree is covered with a plastic film and a shading net, and a growth promoting solution and plant extract are used for watering and spraying.

The invention discloses an inhibition method for induced differentiation of hair follicle stem cells into vascular endothelial cells, comprising: (1), isolating rat hair follicle stem cell; (2), culturing the rat hair follicle stem cells; (3), purifying the rat hair follicle stem cells; (4), inhibiting the induced differentiation of the rat hair follicle stem cells into vascular endothelial cells. In the inhibition method provided herein, vascular endothelial cell growth factor 165 is used for the first time as an inducing factor so that the rat hair follicle stem cells efficiently and directionally differentiate into vascular endothelial cells; the formation and growth of vascular endothelial cells generated by induced differentiation of the hair follicle stem cells can be adjusted, an effective healing of a wound is promoted; it is also possible to provide seed cell sources for solving the problems in vascularization of tissue engineering skin, cell-transplant therapy of ischemic disease and the like.

The process of separating endothelial ancestral cell from human fat tissue relates to method of obtaining seed cell in tissue engineering. Morphological change and cellular immunofluorescene show that the induced cell possesses the phenotype of mature endothelial cell and function of ingesting low density lipoprotein. The detection result with flow cytometer shows that the inducing system possesses very high inducing efficiency. The induced cell forms dendrite bifurcation structure during 3D culture.

The invention discloses a preparation method and application of a silk bracket, and a three-phase silk ligament graft and a preparation method thereof. Silks are woven into a net-shaped bracket with a macroporous structure; the bracket is divided into three areas: a ligament area, a cartilage area and a bone area; mesenchymal stem cells, mesenchymal stem cells carrying with TGF-beta gene lentiviral transfection and mesenchymal stem cells carrying with BMP-2 gene lentiviral transfection are respectively planted in the three areas; the net-shaped bracket modified by the divided areas is rolled to form a bionic ligament graft with a physiological transition structure. The method avoids the problem of stress concentration due to direct connection of the soft and hard tissues; the planting gene-modified mesenchymal stem cells have excellent directional differentiation features, so that the cultivating time and steps of planting a plurality of different cells are saved, and the problem of weak biological fixation of the ligament graft-bone combination part in the existing single-phase tissue engineering ligament and the problem of the traditional method incapable of enabling the cells to grow deeply are solved.

The invention relates to a method for inducing in vitro directional differentiation of stem cells by cell co-culture. An annular gasket is arranged on a bottom of a culture dish. Firstly, stem cells are inoculated in an annular zone and are cultured in an incubator. When the stem cells attach a culture dish wall, a mixed solution containing agarose and sodium alginate is added to the annular zone. When the mixed solution is solidified, a calcium chloride solution is added for a carrying out a calcification reaction and a chitosan is added for carrying out a reaction to form an agarose/sodium alginate/chitosan flat composite gel layer on the stem cells. Inductive cells are inoculated on a surface of the composite gel. By means of the co-culture of the inductive cells and the stem cells, interactions among in vivo cells and between cells and soluble factors can be simulated. By means of the polysaccharide-based agarose/sodium alginate/chitosan flat composite gel, an in vivo extracellular matrix can be simulated. The directional differentiation of the stem cells can be adjusted and controlled through rigidity and thickness of the composite gel. Meanwhile, isolated co-culture of the stem cells and the inductive cells can be achieved so that differentiated cells are easy to collect. The method has an important effect in application of regenerative medicine.

The invention provides a method for fast separating and extracting human Umbilical Cord mesenchymal stem cells (hUC-MSC) from umbilical cord outer layer amnion tissue. The method includes the following steps that fresh collected umbilical cord tissue of a healthy newborn is taken, transported on ice in double-antibody-containing umbilical cord storing transporting liquid, washed and disinfected with 75% ethyl alcohol and normal saline, outer layer amnions are separated bluntly after blood vessels are rejected, mechanically smashed and treated with erythrocyte lysate for 3 min, IV type collagenase digestion is used, and a digestion product is cultured in a suspension mode with a serum-free culture medium after being screened through a 100-200-mesh sieve; medium change is carried out every three to five days, supernatant is taken to detect cell contamination, after the attachment rate in a plate reaches 30-70% and trypsinization is used, cell passage amplification is collected centrifugally, and confluence is carried out when the cell confluence rate reaches 90% or above, the cells are collected for cryopreservation, and the biological characteristics of hUC-MSC are detected.