181 results about "Stem Cell Isolation" patented technology

A method of augmenting and / or repairing an intervertebral disc by administering stem cell material into the disc. The stem cells may be undifferentiated cells, or they may be cells that have differentiated and have subsequently been dedifferentiated. The stem cells may be induced to express at least one characteristic of human intervertebral disc cells, such as fibroblast cells, chondrocyte cells, or notochordal cells, by exposing them to agents and / or environments calculated to induce the desired differentiation. In some embodiments, the stem cell material may be provided in conjunction with a collagen-based material, which may be a collagen-rich lattice or particles of collagen material. The stem cell material may be provided as a stem cell isolate, which may be substantially free of non-stem cell material. Other therapeutic agents may be administered with the stem cell material.

The present disclosure describes mammalian pluripotent embryonic-like stem cells (ELSCs) isolated from corneal limbal tissue, a non-embryonic tissue. The ELSCs of the present disclosure are capable of proliferating in an in vitro culture, maintain the potential to differentiate into cells of endoderm, mesoderm, and ectoderm lineage in culture, and are capable of forming embryoid-like bodies when placed in suspension culture. Thus, these cells possess multi-lineage differentiation potential and self-renewing capability. ELSCs may be a promising therapeutic tool, and may provide new therapeutic alternatives for various diseases, conditions, and injuries.

Methods are presented in which release of stem cells from skeletal muscle is quantitated and correlated with severity of a disease or trauma, a future treatment option, prognosis, and / or anticipated time to recovery. Most preferably, the stem cell is a BLSC and / or an ELSC, and the stem cell isolation for the cell count is performed using sedimentation or filtration as principal separation step, thereby avoiding commonly used complicated, expensive, and time-consuming processes such as antibody-based separation and fluorescence-activated cell sorting.

The present invention has its object to provide a material for separating stem cell and a filter for separating stem cell, each is capable of selectively separating and recovering, in a simple and easy manner, stem cells from body fluids or biological tissue-derived treated fluids, a method for separating and recovering stem cells, and stem cells obtained by such method. The present invention is a material for separating stem cell which has a density K of 1.0×104≦K≦1.0×106 and a fiber diameter of 3 to 40 μm; a filter for separating stem cell which comprises the material for separating stem cell as packed in a container having a fluid inlet port and a fluid outlet port; a method of separating and recovering stem cells which comprises using the material for separating stem cell or the filter for separating stem cell; and a method of producing a multipotent cell fraction.

The present invention relates to a reagent kit of the isolation of a bone marrow umbilical cord blood stem cell in vitro and the application method thereof. The present invention is characterized in that Lin antibody red blood cell precipitant cell preserving solution, and sodium iothalamate-polysucrose 400 # or HISTOPAQUE1077 or Ficoll are contained in the reagent kit, and each reagent solution is preserved dividually. The present invention adopts the method which combines a negative collection method and a density method, thus the surface of the stem cell which is collected and abstracted has no any label, the cells which are isolated and abstracted are Lin antigenic negative cells, the method is simple, and the present invention can be directly used to isolate and abstract the stem cells clinically. The reagent kit of the present invention can operate the industrialized production, to realize the promotion of the isolation and purification technology of the human hematopoietic stem cell, leading the doctor to conveniently cure the patient with the stem cells like the medicine usage.

The invention includes compositions of stem and progenitor cells recovered from bone marrow or cord blood containing most of the viable CD34+ cells and substantially depleted of red blood cells resident in the original sample, without any xenobiotic additives to aid cell separation. The invention also includes a system and method for preparing the compositions. The system includes a bag set and a processing device, which utilizes an optical sensor, microcontroller, servo motor, accelerometer, load cell, and battery. The system and method utilize centrifugation to stratify the cells into layers and then separate and transfer the stem cells into a stem cell bag. The processing device's microcontroller receives input from the device's accelerometer, load cell and optical sensor to direct the metering valve in the bag set to open and close to permit the transfer of as many stems cells as possible with as few red cells as possible.

The invention discloses a separation and culture method using ginseng cambium stem cells. The separation and culture method using ginseng cambium stem cells is characterized by comprising the following steps: (1) firstly performing disinfection treatment on the root tubers with the diameter of 2 cm of northeast ginseng by 0.1% mercury bichloride for 10 minutes, and then transversely cutting the northeast ginseng into semicircular slices with the thickness of 0.2 cm and containing periderms, phloems and cambiums; (2) placing the semicircular slices on a WPM (woody plant medium) containing 0.1-4.0 mg / L of picloram and 0.1-4.0 mg / L of 2,4-dichlorphenoxyacetic acid, and culturing at 25 DEG C in the dark; (3) taking out the obviously-multiplied explants of the cambiums after two weeks, separating the cambium stem cells, transferring the cambium stem cells on a subculture medium and culturing, wherein the subculture medium is a WPM containing 0.1-4.0 mg / L of picloram and 0.1-4.0 mg / L of 2,4-dichlorphenoxyacetic acid, and subculture is performed every two weeks. According to the method, separation and culture for ginseng stem cells are performed by using the unsplit cambium stem cells, and the cambium stem cells are infinitely multiplied, so that production can be effectively expanded.

The present invention relates to novel miRNA molecules, more particularly to novel miRNA molecules isolated from human embryonic stem cells. The miRNA molecules provided by the present invention can be usefully used as a molecular marker for early developmental stages of undifferentiated human embryonic stem cells. Also, the miRNA molecules of the present invention may play an important role in the regulation of mammalian embryonic stem cells. Therefore, the miRNA molecules can be usefully used for analyzing regulatory networks of human embryonic stem cells.

The invention develops a simplified method for the in-vitro isolation and expansion of umbilical mesenchymal stem cells (MSCs) and confirms that the MSCs obtained by the method have the effects of immunoregulation and inflammation inhibition in the treatment of rheumatoid arthritis.

The present invention relates to stem cell separating and purifying technology, and is especially one kind of stem cell separating liquid and its usage in separating stem cell. The stem cell separating liquid consists of meglucamine diatrizoate 0-10 weight portions, methylol cellulose 0-10 weight portions, cesium chloride 0-1.0 weight portions, and sucrose polymer 0-10 weight portions. It is prepared through mixing the said ingredients and adding 0.01 mol / l concentration phosphate buffering liquid. When it is applied, it is diluted with 0.01 mol / l concentration phosphate buffering liquid to density of 1.080-1.120 g / ml. The present invention has high stem cell separating effect.

The invention provides a novel menstrual blood-derived mesenchymal stem cell separation method, belonging to the field of methods for separating stem cells from menstrual blood. In order to solve the problems of long centrifugal time, relatively serious cell injury, low cell yield and the like existing in a lymph separating medium method in the traditional method, the invention provides the novel menstrual blood-derived mesenchymal stem cell separation method comprising the steps of with source-wide menstrual blood as a material, storing by using a special preserving fluid in a collecting process; then, carrying out primary separation by using a density gradient centrifugation method of a lymphocyte separation tube, and optimizing the centrifugal time and centrifugal rotating speed in separation; and next, carrying out secondary separation according to the wall attachment growth characteristic of the stem cells, simultaneously separating and amplifying the stem cells, and maintaining the activity of the stem cells to the maximum extent in the separation process. The obtained menstrual blood-derived mesenchymal stem cell is high in purity, quantity and application value. The novel menstrual blood-derived mesenchymal stem cell separation method is simple in operation and low in cost.

The invention relates to a method for cryopreservation and resuscitation of decidua parietalis tissue and isolation and culture of mesenchymal stem cells. The method includes the steps of: (1) pretreatment of placenta tissue; (2) separation and treatment of placenta decidua parietalis tissue; (3) cryopreservation of placenta decidua parietalis tissue; (4) resuscitation of placenta decidua parietalis tissue; (5) separation of mesenchymal stem cells from placenta decidua parietalis tissue; and (6) subculture of decidua parietalis stem cells. The method provided by the invention cuts the stripped decidua parietalis tissue into pieces for direct cryopreservation. Treatment of each tissue and culture of isolated cells both need certain time and staff consumption. Therefore, the practice of cryopreservation of decidua parietalis tissue and resuscitation for stem cell isolation when needed relatively more conforms to cost-effectiveness.

The invention discloses an umbilical cord mesenchymal stem cell (MSC) separation culture and amplification method. The umbilical cord MSC separation culture and amplification method comprises the steps of (1) clearing, (2) treatment, (3) culture, and (4) inoculation and passage. The umbilical cord MSC method obtained by adopting a tissue mass separation method is simple, the cost is low, cell activity is good, purity is high, the final harvested cell concentration is high, the obtained MSC method is simple, MSCs can be quickly and efficiently separated, the culture time is shortened, after culture is conducted for 4-5 days, it can be seen that the cells climb out from tissue masses, the final harvested cell concentration is high, and the entire umbilical cord MSC culture process can be standardized, programmed and normalized.

The invention discloses a kit for mesenchymal stem cell culture and its application. The kit provided by the invention includes a cell culture solution A and a cell culture solution B. The cell culture solution A is a Knockout-DMEM culture medium containing 80-120ng / ml cytokine LIF and 80-120ng / ml cytokine bFGF. The cell culture solution B is fetal bovine serum. The kit provided by the invention can separate umbilical cord mesenchymal stem cells from an in-vitro umbilical cord and culture the umbilical cord mesenchymal stem cells, and has the advantages of rapid cell expansion speed and high expansion efficiency. After multiple subculturing, cell totipotency can still be maintained. In the invention, the problems of umbilical cord mesenchymal stem cell separation purity, quantity and primary culture time are solved, and a lot of umbilical cord mesenchymal stem cells can be obtained. Thus, the kit provided in the invention provides abundant sources for making use of umbilical cord mesenchymal stem cells to conduct further experimental research.

The invention provides a preparation method of placenta-source decidua parietalis mesenchymal stem cells by separation and refrigeration. According to the preparation method, puerpera is screened before placenta acquisition, and placenta from healthy mature puerperas volunteers who give birth to baby boys is finally selected as the acquisition target; separation and amplification are carried out for large-scale culture until the third generation, and refrigeration and preservation are carried out; and an intermediate product obtained from the separation process and refrigeration process undergoes a series of quality detections such as maternal detection, and cells are resurrected and washed for later use after refrigeration. There is no obvious reduction of motility rate, and biological functions of cells are complete. The placenta-source decidua parietalis mesenchymal stem cells have a simple component, and the whole process from raw material acquisition to cell preparation is monitored clinically, unitedly and systematically so as to realize a clinic standard product of mesenchymal stem cells. Besides, decidua parietalis comes from mother and is more suitable to be used by maternal body.

The invention relates to a method for separation and purification, long-term passage cultivation, cryopreservation and recovery of sheep spermatogonia stem cells. The method includes the steps that various solutions are prepared, 2-3 g of tissue blocks of the seminiferous tubule are taken out from the testis of a sheep of 3-4 mouths of age, single-cell suspension is prepared through enzymolysis and digestion of two steps, purified spermatogonia stem cells are separated according to the difference adherence method, the long-term passage cultivation of the sheep spermatogonia stem cells is achieved through an SSCM culture solution, and the sheep spermatogonia stem cells are cryopreserved and recovered after the long-term passage cultivation. By the technical scheme, the sheep spermatogonia stem cells with high purity can be obtained without using specific sheep spermatogonia stem cell antibodies, a long-term passage cultivation system of the sheep spermatogonia stem cells is built, and the method for preserving the sheep spermatogonia stem cells for a long term through a liquid nitrogen freezing method is successfully provided. The sheep spermatogonia stem cells achieving separation, purification and passage according to the method are cultivated at least for more than 40 generations, and the longest cultivation period reaches four years if the cryopreservation time is included.

The invention provides a method for establishing a uterine membrane stem cell bank. The method comprises the steps of menstrual blood collection, uterine membrane stem cell separation and degerming, amplification, analysis, detection, long-term storage and regular detection. The invention provides a degerming method which is simple and easy to operate. The method provided by the invention can obtain a large quantity of higher-purity uterine membrane stem cells by carrying out amplification culture on the uterine membrane stem cells with substances (such as bacteria) removed and can be used for storing the uterine membrane stem cells for a long time, thereby providing a large quantity of uterine membrane stem cell resources for future clinical and scientific research application study.

The invention provides a kit for culture of adipose-derived stem cells, wherein the kit includes a kit body, a plurality of reagent bottles and a plurality of ice bags; the plurality of reagent bottles are internally filled with a tissue preservation liquid, a basal culture medium, a culture medium additive, a growth factor I, a growth factor II, a tissue decomposition liquid, a cell digestion liquid, a washing liquid and a frozen stock liquid respectively. The kit provided by the invention can separate the adipose-derived stem cells from adipose tissues, and cultures and preserves the adipose-derived stem cells, solves the problems of small separation number and low purity of the adipose-derived stem cells, low culture survival rate, low culture efficiency, nonideal frozen resurrection rate and the like, makes the adipose-derived stem cells subjected to multi-generation amplification and not generate differentiation in an ideal nutrient equilibrium state, provides a rich source for further experimental research with use of the adipose-derived stem cells, and is quite suitable for culture of the adipose-derived stem cells.

The present invention for the first time demonstrated that:(1) bone marrow-derived pluripotent tissue stem cells can be induced in peripheral blood by intravenously administering tissue extract prepared from isolated skin pieces;(2) the substance in the isolated skin pieces, which is responsible for mobilizing bone marrow-derived pluripotent tissue stem cells to peripheral blood, is HMGB1; and(3) HMGB1 with the activity of mobilizing bone marrow-derived pluripotent stem cells to peripheral blood can be easily purified from cultured cells.

The invention discloses an isolated culture and identification method of porcine adipose-derived stem cells. The method comprises the following steps: 1) isolated culture and purification of adipose-derived stem cells; and 2) subculturing of adipose-derived stem cells. The adipose-derived stem cells obtained by the method disclosed by the invention have good genetic stability, and the genetic stability is still kept after the adipose-derived stem cells are continuously cultured in vitro to the 15th generation; with good dryness, the adipose-derived stem cells still have the surface antigen characteristics of mesenchymal stem cells after cultured in vitro to the 18th generation; and with good multi-directional differentiation property, the adipose-derived stem cells have the ability of differentiating into osteoblasts, chondroblasts and lipoblasts.

The invention relates to the technical field of adipose-derived stem cell isolated culture, and discloses a dental pulp stem cell isolated culture method. The dental pulp stem cell isolated culture method comprises the following steps: washing adipose tissues, digesting the adipose tissues by using collagenase Type I, separating adipose stromal vascular fractions, then carrying out scavenging-precipitation, planting and culturing obtained primary cells to obtain the primary cells in the end, and carrying out primary cell passage. The dental pulp stem cell isolated culture method has the beneficial effects that by standardizing the dental pulp stem cell isolated culture method and various parameters in concrete operations, the adipose-derived stem cell isolated culture efficiency is improved, the output rate is increased, and the production cost is reduced.

The invention relates to a canine stem cell isolated culture technology, and especially relates to a method for in vitro simultaneous insolated culture of canine bone marrow mesenchymal stem cells and multifunctional hematopoietic stem cells. The method is characterized in that adherent culture of primary mesenchymal stem cells is carried out based on the separation of bone marrow mononuclear cells, half medium change is adopted, and a mononuclear cell culture suspension is added, that is, hematopoiesis microenvironment provided by the adherent-growth mesenchymal stem cells is used to culture the suspending-growth multifunctional hematopoietic stem cells, so the achieve the simultaneous isolated culture of the canine bone marrow mesenchymal stem cells and the multifunctional hematopoietic stem cells is realized.

The invention relates to an SPA-antibody tripolymer, a cell treating kit containing the tripolymer, a preparation method and application thereof. The tripolymer comprises an anti erythrocyte antibody, an SPA and an antibody of a certain anti leukocyte antigen or other antigens. The tripolymer is combined with an erythrocyte through the anti erythrocyte antibody per se, the other antibody is combined with a corresponding leukocyte antigen (or other antigens), and then leukocyte (other cells, factors) and the erythrocyte are deposited through an erythrocyte sedimentation process or other methods of density gradient centrifugation and the like, thereby achieving the purpose of eliminating ingredients of corresponding cells and the like, connecting the erythrocyte with a corresponding antigen by the SPA, and being used as a mean for detecting a certain antigen. The invention has convenience and simpleness, and can be directly used for clinically separating and extracting stem cells / progenitor cells; and the collected and extracted stem cells / progenitor cells have no external markers. Meanwhile, the kit based on the tripolymer can be industrially produced so as to realize the popularization of the separation and purification technology of hematopoietic stem cells.

The invention provides a method for preparing a placenta hematopoietic stem cell preparation. The method comprises the following steps: (1) digesting placenta tissues with an enzyme solution so as to obtain a digestive product and removing tissue residues in the digestive product to obtain enzymatic hydrolysate; (2) mixing the enzymatic hydrolysate with a hydroxyethyl starch solution, and layering the mixed materials to obtain a nuclear cell suspension; and (3) mixing the nuclear cell suspension with a frozen stock solution, wherein the enzyme solution contains type I collagenase, DNA enzyme I, dispase and hyaluronidase, and the frozen stock solution contains dimethyl sulfoxide, hydroxyethyl starch, human albumin and dextran. According to the technical scheme, the separation efficiency of the placenta hematopoietic stem cell can be improved; and the frozen placenta hematopoietic stem cell can be directly applied to stem cell treatment when heated to a room temperature.

The invention provides a method for separating and culturing human umbilical cord mesenchymal stem cells, which comprises the following steps: taking umbilical cord of fetus born after the normal period of gestation by caesarean section, flushing by PBS under aseptic condition, removing the bloodstain; using an aseptic scissor in an animal cell medium to cut the washed umbilical cord to tissue blocks, removing blood vessel and crushing, then fully mixing with the animal cell medium and oscillating, centrifuging, mixing the centrifuged deposition and a MesenCult-XF complete medium and oscillating, and uniformly planting in a cell culture dish, placing in an incubator with the temperature of 37 DEG C and 5% of CO2 for culturing for 5 days, sucking the tissue blocks out and discharging, refilling the MesenCult-XF complete medium, and replacing the medium every three days for continuously culturing to obtain the umbilical cord mesenchymal stem cells. According to the invention, the animal serum is used, the operation is simple, the repeatability is good, the security is high, and the obtained cell enables stabilization through multitime passage.

The invention discloses umbilical cord blood mesenchymal stem cell separation liquid and an umbilical cord blood mesenchymal stem cell separation flow. The separation liquid is prepared by mixing saccharosan of which the concentration is 9 percent and meglumine diatrizoate of which the concentration is 33.9 percent in the ratio of 26.88:10, wherein the concentration of the separation liquid is 1.073+ / -0.01g / L. A two-step method for separating umbilical cord blood mesenchymal stem cells comprises the following steps of: 1, precipitating red cells by a hydroxyethyl starch; and 2, separating the mesenchymal stem cells by the separation liquid of which the concentration is1.073g / L. The mesenchymal stem cell with the highest proportion in umbilical cord blood can be obtained by the separation liquid and the separation flow, the proportions of lymphocytes and mononuclear cells are very low, and the obtained cells are good in state and high in activity. The used materials and reagents are medical products, are non-toxic, do not have heat sources, can be industrially produced and are convenient to store. The used method is an optimized flow, is high in stability and repeatability, can be also used for separating human marrow and peripheral blood mesenchymal stem cells, is the best method for separating and extracting the mesenchymal stem cells in the current density gradient centrifugation and is better than other products and methods.

The invention discloses a method for obtaining stem cell by using serum of the same cord blood, belonging to the field of biotechnology and tissue engineering. Serum of the same cord blood is used to replace fetal calf serum, no cell factor or stroma cell support is added, and the strategy of combining a microcarrier and a bioreactor is adopted, thus realizing co-culture of cord blood source hemopoietic stem cell and mesenchymal stem cell. The method of the invention can support fine augmentation of UCB-HSCs and UCB-MSCs under three-dimensional conditions, completely replace the role of fetal calf serum and avoid immunological rejection among variants; cytodex-s microcarrier can provide adhesion surface for UCB-MSCs with adherence growth characteristics, so that the UCB-MSCs can realize possible adherence suspension culture in a three-dimensional dynamic environment; the natural settling method is adopted to separate and obtain the suspending UCB-HSCs and the UCB-MSCs which adheres to the microcarrier.

The present invention features stems cells from lungs, methods of isolating such stem cells, methods of diagnosing lung diseases, and method of treating lung diseases, including cancer, chronic obstructive pulmonary disease, cystic fibrosis, and emphysema.

The invention discloses a biological scalp and hair follicle nourishing and hair growing essence and a preparation method thereof. The essence is added with a human skin stem cell factor nano liposome-exosome complex cultured and prepared by using a human skin stem cell isolation, amplification and culture technique, and supplemented with a Chinese herbal medicinal extract essence to organically combine scalp, hair follicles and hairy roots, so that the growth environment of hair is fundamentally improved, the scalp cell activity and density are improved, hair transformation is promoted, the scalp cell activity and density are improved and the functions of increasing the hair, nourishing the hair and caring the hair are achieved.

The invention belongs to the field of stem cell culture, and particularly relates to a cell culture medium and a stem cell culture method. The invention provides a stem cell culture medium. The stem cell culture medium comprises a basic culture medium, a platelet lysate, an L-Glutamine and a mitomycin c. The invention also provides a stem cell separating method. The stem cell separating method comprises the following steps that the in-vitro tissue and the stem cell culture medium are mixed to be placed in a culture flask for culturing until cells creep out from the in-vitro tissue, the in-vitro tissue is scraped; the culture of the cells is continued, when the fusion degree of stem cells in the culture flask reaches 80% to 90%, the cells are passage-cultured after digestion. The stem cell culture medium can avoid the risk that the existing stem cell culture medium contains mostly animal serum so as to introduce exogenous virus; the stem cell separating method is simple to operate.