57 results about "Insulin Secreting Cell" patented technology

Methods and compositions are described for the treatment of type I insulin-dependent diabetes mellitus and other conditions using newly identified stem cells that are capable of differentiation into a variety of pancreatic islet cells, including insulin-producing beta cells, as well as hepatocytes. Nestin has been identified as a molecular marker for pancreatic stem cells, while cytokeratin-19 serves as a marker for a distinct class of islet ductal cells. Methods are described whereby nestin-positive stem cells can be isolated from pancreatic islets and cultured to obtain further stem cells or pseudo-islet like structures. Methods for ex vivo differentiation of the pancreatic stem cells are disclosed. Methods are described whereby pancreatic stem cells can be isolated, expanded, and transplanted into a patient in need thereof, either allogeneically, isogeneically or xenogenically, to provide replacement for lost or damaged insulin-secreting cells or other cells.

The invention relates to methods for generating insulin secreting cells from precursor stem cells or from adult pancreatic exocrine cells. The methods of the invention are useful, for example, for generation of glucose sensitive insulin-secreting beta-cells suitable for transplantation, as well as for in situ development of insulin-secreting cells in a patient in need thereof. Further, the method of the invention relates to methods for preventing premature differentiation of precursor stem cells into insulin-secreting beta-cells. Still further, the invention relates to assay methods for identification of compounds that prevent or activate beta-cell differentiation.

The invention discloses an induction culture medium which comprises a solvent DMEM (dulbecco's modified eagle medium)-high sugar culture medium, platelets, all-trans vitamin A acid, activin A, glicetin I, a fibroblast growth factor, beta cytosine, an insuline-like growth factor, nicotinamide, polypeptide, pentagastrin and mercaptoethanol. Physical oscillation and collagenase / pancreatin dual digestion are performed to efficiently separate fat mesenchymal stem cells from human fat tissues. The fat mesenchymal stem cells are cultured for proliferation and subcultivation, and the induction culture medium is utilized to induce the fat mesenchymal stem cells to become insulin secreting cells, so that more than 40% of the fat stem cells are induced to differentiate into the insulin secreting cells.

Aspects of the disclosure generally relate to insulin delivery systems and compositions having a insulin secreting β cell line or insulin secreting recombinant non-β cells sequestered in a glucose-responsive material. The disclosed insulin delivery systems can be surgically implanted in a host and can provide a continuous source of insulin from the cells contained in the device. The combination of living cells with a glucose-responsive material provides a hybrid insulin delivery system that delivers physiologically relevant amounts of insulin in response to physiologically relevant glucose levels. In some aspects, the disclosed devices have a biphasic release of insulin in response to sudden increases in glucose concentrations in the fluids bathing the device.

Proliferating pancreatic islet cells obtained by the method of isolating a population of cells that preferably includes predominantly islet precursor cells that express one or more marker associated with an islet precursor cell and providing the precursor cells with one or more a pancreatic differentiation agent so that a population of cells is obtained that has a high proportion of cells with phenotypic characteristics of functional pancreatic islet β-cells. Optionally, the precursor cells are pretreated by providing them with one or more cell expansion agent to increase the number of cells in the population prior to differentiation. The pancreatic differentiation agent composition comprises a gastrin / CCK receptor ligand, e.g., a gastrin, in an amount sufficient to effect differentiation of pancreatic islet precursor cells to mature insulin-secreting cells. The cell expansion agent composition comprises one or more epidermal growth factor (EGF) receptor ligand in an amount sufficient to stimulate proliferation of the precursor cells. The methods of treatment include transplanting either undifferentiated precursor cells and providing the pancreatic differentiation agent either alone or in combination with the cell expansion agent in situ, or transplanting the functional pancreatic islet β-cells into the patient. The pancreatic islet β-cells can be used for drug screening, and replenishing pancreatic function in the context of clinical treatment.

The invention discloses a method for constructing tissue engineering islet by adipose derived mesenchymal stem cells and the obtained tissue engineering islet. The method employs adipose derived mesenchymal stem cells with extensive sources to perform culture on islet acellular matrix, so that the acellular matrix can provide a three-dimensional space suitable for growth to adipose derived mesenchymal stem cells so as to promote cell adhesion, proliferation and differentiation, and formation of a three-dimensional structure engineering islet tissue able to stably secrete insulin. The method realizes gathering of insulin secreting cells, enhances cell utilization and is expected to promote the effect of repairing islet injury by tissue engineering method.

The invention belongs to the field of biomedicine, and relates to an inducer for inducing human adipose derived stromal cells into insulin-secretion cells, an induction medium and a method. The inducer for inducing human adipose derived stromal cells into insulin-secretion cells consists of the following components: a rhodiola rosea injection, nicotinamide, taurine and GLP-1. Every 1,000ml of the induction medium containing the inducer comprises 20-40g of the rhodiola rosea injection, 0.97-1.47g of the nicotinamide, 0.25-0.50g of the taurine, 26.84-40.27mg of the GLP-1 and the balance being a human adipose derived stromal cell serum-free medium, which are mixed uniformly, filtered and degermed. According to the inducer for inducing human adipose derived stromal cells into insulin-secretion cells, the induction medium and the method, differentiation of the human adipose derived stromal cells into the insulin-secretion cells is induced by using the injection of a traditional Chinese medicine, namely, rhodiola rosea, and each component is safe and nontoxic. The method has few steps and is short in time and high in the inducing efficiency.

Methods for increasing glucose induced insulin secretion in an insulin secreting cell by inducing senescence in the cell are provided. Further, methods for treating diabetes, by providing cells with increased glucose induced insulin secretion to a subject, as well as a population of modified insulin secreting cells are provided.

The invention discloses a medium composition for inducing mesenchymal stem cells into insulin-secreting cells. The medium composition comprises a medium A, a medium B and a medium C. The medium A comprises the following solutes: a fetal calf serum with a concentration of 4.75 to 5.25 ml / L and activin A with a concentration of 4.75 to 5.25ng / mL. The medium B comprises the following solutes: retinoic acid with a concentration of 0.95 * 10<-5> to 1.05 * 10<-5> mol / L, EGF with a concentration of 19 to 21ng / ml, bFGF with a concentration of 19 to 21ng / ml, glutamine with a concentration of 1.9 to 2.1 mmol / L, 0.95 to 1.05% of non-essential amino acids and 1.9 to 2.1% of B27. The medium C comprises the following solutes: a fetal bovine serum with a concentration of 4.5 to 5.5 ml / L, exendin-4 with a concentration of 19 to 21 ng / ml, activin A with a concentration of 9.5 to 10.5 ng / ml, nicotinamide with a concentration of 9.5 to 10.5 mmol / L, 1.9 to 2.1% of B27 and 0.95 to 1.05% of N2. An induction culture method provided by the invention is applicable to a variety of tissue-derived mesenchymal stem cells and has the advantages of no introduction of viruses, easy operation and high efficiency.

The invention relates to a method for inducing placenta-derived mesenchymal stem cells to be differentiated into islet-like cells. According to the method, placenta-derived mesenchymal stem cells are induced to be differentiated to islet-like cells and are identified by using cytokines, such as basic fibroblast growth factors (bFGF), hexadecadrol and beta-mercaptoethanol. The method lies in further investigating the differentiation potential of placenta-derived mesenchymal stem cells, discussing and studying optimum conditions of in-vitro separation, culture and differentiation of the placenta-derived mesenchymal stem cells, studying the possibility and the feasibility of inducing the placenta-derived mesenchymal stem cells to be differentiated into the islet-like cells, seeking an optimum inducing system of inducing differentiation in vitro and further finding a method for inducing the placenta-derived mesenchymal stem cells to be differentiated to insulin-producing cells (IPCs). Therefore, the method provides a foundational research basis for clinical application of the placenta-derived mesenchymal stem cells to the insulin-producing cells.

The invention provides a stem cell preparation for treating diabetes mellitus. The stem cell preparation comprises: stem cells separated and extracted from human blood or tissues and organs and / or islet-like cells induced and differentiated from the stem cells and a cell preservation solution containing 0.5% of heparin calcium, wherein the cell content is 1*10<5> to 5*10<7> / ml. The invention also discloses a preparation method of the stem cell preparation. The method comprises the following steps: (1) separation and purification of the stem cell; (2) in vitro culture of the stem cells or in vitro induction and differentiation of the stem cells to form islet-like cells; (3) diluting the cell obtained in the step (2) by using the cell preservation solution to prepare a stem cell preparation; (4) quality detection of the stem cell preparation. The stem preparation is transplanted into the body and can be differentiated into insulin-secreting cells to secret insulin, and the blood sugar concentration of a person with diabetes is effectively reduced. Thus, a new way is provided for overcoming treatment of the diabetes dependent on the insulin and curing the diabetes once and for all.

The invention discloses a method for the in vitro induction of hAMSCs differentiated into ISCs. According to the method, the extracted hAMSCs are cultured for 7 days by a serum-free high sugar-DMEM induction medium containing niacinamide of 10mmol / L and an N2 supplement. The hAMSCs induced by the method of the invention express insulin gene mRNA and PDX-1 gene mRNA, and the content of insulin in the supernatant of the culture reaches above 330muIU / ml; and the method of the invention allows the in vitro induction of the hAMSCs differentiated into the ISCs to be realized.

The invention discloses a method for directionally inducing insulin-secreting cells by endometrial stem cells, which comprises the following steps of: 1, separation culture and amplification of the endometrial stem cells; and 2, in vitro inducing differentiation. The step of in vitro inducing differentiation comprises the following specific steps of: digesting the fourth to sixth generation of cells with good growth conditions, which are obtained in the step 1, by pancreatic enzymes; when the cells are observed under a microscope to be changed into single circles, stopping digesting by a DMEM(dulbecco's modified eagle medium) culture medium containing fetal calf serum with the volume concentration of 13 to 17 percent; centrifuging and washing the obtained cell precipitates by a PBS (Phosphate Buffer Solution); adding an inducing culture medium into the cells washed by the PBS; placing the obtained cells into an incubator with the concentration of carbon dioxide CO2 of 4 to 6 percent,the saturated humidity of 94 to 96 percent and the temperature of 36 to 38 DEG C for culturing; and culturing the cells for 13 to 15 days to obtain the insulin-secreting cells. The obtained insulin-secreting cells are used for treating diabetes mellitus so as to provide a novel seed resource for stem cell therapy.

The invention discloses a method for constructing and differentiating a pancreatic stem cell line from human insulin. The pancreatic stem cell line is a pancreatic stem cell amplified from human insulin mass and has a multi-lineage differentiation potential. The induced pancreatic stem cell line can be differentiated to form nerve-like cells, fat-like cells, bone-like cells and functional insulin-like cell mass. The functional insulin-like cell mass is transplanted to a body of a nude mouse model suffered from diabetes, so that the blood sugar concentration of the nude mouse model can be reduced and normal blood sugar level can be maintained within one month.

The invention relates to the field of biomedicine, in particular to a method for inducing human mesenchymal stem cells to differentiate into insulin-secreting cells in vitro, which includes, according to the technical scheme, constructing recombinant eukaryotic expression plasmids of SRF (serum response factor) gene; preliminarily inducing human mesenchymal stem cells into nestin cells first; further inducing the nestin cells into pancreatic progenitor cells; transfecting an eukaryotic expression vector containing the SRF gene to the cells above; and finally inducing the cells into insulin-secreting cells, wherein the eukaryotic expression vector comprises pEGFP (plasmid enhanced green florescence protein), pcDNA or pCMV. By the method, new seed cell sources for cell therapy of diabetes mellitus are provided, new theoretical and experimental basis is provided for directed induction of mesenchymal stem cells differentiating into insulin-secreting cells and clinical application of the insulin-secreting cells, and new models for drug development and screening related to the insulin-secreting cells are provided.

Methods and compositions are described for the treatment of type I insulin-dependent diabetes mellitus and other conditions using newly identified stem cells that are capable of differentiation into a variety of pancreatic islet cells, including insulin-producing beta cells, as well as hepatocytes. Nestin and ABCG2 have been identified as molecular markers for pancreatic stem cells, while cytokeratin-19 serves as a marker for a distinct class of islet ductal cells. Methods are described whereby nestin and / or ABCG2-positive stem cells can be isolated from pancreatic islets and cultured to obtain further stem cells or pseudo-islet like structures. Methods for ex vivo differentiation of the pancreatic stem cells are disclosed. Methods are described whereby pancreatic stem cells can be isolated, expanded, and transplanted into a patient in need thereof, either allogeneically, isogeneically or xenogenically, to provide replacement for lost or damaged insulin-secreting cells or other cells.

The invention provides a method for induced differentiation of human amniotic mesenchyme cells in vitro into insulin-secreting cells. The steps include: isolation culture of primary cells; purification of primary cells; primary cell identification by a flow cytometer; 2-mercaptoethanol, B27, bFGF, and nicotinamide induction; and cell morphology and function identification after induction. The method for induced differentiation of human amniotic mesenchyme cells in vitro into insulin-secreting cells provided by the invention induces human amniotic mesenchymal cells to differentiate into nestin positive cells, induces nestin positive cell amplification and differentiation into precursor cells, and induces insulin-secreting cell maturation, confirms that amniotic mesenchyme cells have the possibility of transformation into insulin-secreting cells in specific environment, amniotic mesenchyme cells have wide sources, and no ethical obstacle exists, therefore the method provides a new idea for clinical stem cell transplantation.

The invention discloses an artificial islet or artificial pancreas and a preparation method thereof. The artificial islet or artificial pancreas includes a supporting body with a porous structure andcells capable of secreting insulin and growing in the porous structure of the supporting body. The preparation method of the artificial islet or artificial pancreas comprises the steps that a PLGA porous sphere supporting body is prepared; the cells capable of secreting the insulin are planted in the PLGA porous sphere supporting body, and the cells planted in the PLGA porous sphere supporting body are detected. The construction of the artificial islet or artificial pancreas is based on the porous structure of PLGA or a polylactic acid polymer sphere and the good biodegradability and biocompatibility thereof, and effectively simulating the structure and physiological function of an islet is facilitated. The insulin secretory cells are set in the constructed artificial islet or artificial pancreas porous sphere, the cell morphology is good, the overall morphology and size are similar to normal mouse islets, and an effective solution is provided for solving the shortage of donors in clinical islet transplantation.

A pharmaceutical composition for preventing or treating steroid-induced diabetes (SID) and a health functional food, and more particularly to a pharmaceutical composition for preventing or treating SID and a health functional food, contain a Ceriporia lacerata culture extract as an active ingredient. The pharmaceutical composition and the health functional composition have the effect of protecting INS-1 insulin-secreting cells by blocking the steroid-mediated proliferation inhibition and apoptosis induction of INS-1 insulin-secreting cells. Thus, the pharmaceutical composition and the health functional composition can be developed as the active ingredient of pharmaceutical or functional food compositions serving to protect or regenerate β-cells by blocking the steroid-mediated growth inhibition and apoptosis of β-cells.

The invention discloses a method for induced differentiation of amnion epithelial stem cells into pancreatic beta cells with biological functions. The method comprises the steps as follows: 1) separation, culture, amplification and identification of amnion epithelial stem cells; 2) induced differentiation in vitro: the second-third generation of amnion epithelial stem cells are selected for experiments and subjected to induced differentiation by a culture medium containing nicotinamide. Insulin secretion cells which can express specific markers of pancreatic beta cells and have functions of normal pancreatic beta cells are obtained by 14 days of differentiation in vitro; and 3) in vivo transplantation: insulin secretion cells produced by induction are transplanted into type-1 diabetes miceand can obviously relive the hyperglycemia state of mice.

The invention provides a culture medium for separating and inducing human adipose stem cells into pancreatic beta cells and a use method of the culture medium. The optimal culture medium is designed; gene transfection is not required, so that the risk of gene modification and cancers is avoided, few induction steps are used, the use is simple, and the induction time is short and is shortened by 30% in comparison with the prior art; the culture medium is safe, nontoxic and high in success rate of induction. Moreover, after autogenous adipose mesenchymal stem cells are induced and differentiated into insulin-secreting cells, rejection and ethical problems are avoided after autotransplantation, and the safety is high, so that the clinical application prospect is wide. The culture medium is simple, feasible, low in cost and good in use effect.

Process for preserving insulin secreting cells intended to be transplanted, including the following steps: introducing an initial volume of culture medium and insulin-secreting cells into a culture container, providing a culture medium source, and replacing, at time intervals below 8 hours, the culture medium in the culture container with culture medium from the source so as to renew the culture medium contained in the container.

Disclosed is a method for differentiating human adult stem cells into insulin-secreting cells. Human adult stem cells, isolated from the subcutaneous adipose tissues around the eyes, are induced to differentiate into insulin-secreting cells in a medium in the presence of cytokines and growth factors including B27 supplement, fibroblast growth factor-2, epidermal growth factor, nicotinamide, glucagon-like peptide-1, activin A, insulin-like growth factor, betacellulin, etc., with a glucose shift from a high concentration to a low concentration. Having the ability to producing insulin and C-peptide at high levels, the insulin-secreting cells can be excellent curatives for type 1 diabetes mellitus.