44 results about "Stem cell biology" patented technology

The present invention relates to the field of stem cell biology, in particular the linage specific differentiation of pluripotent or multipotent stem cells, which can include, but is not limited to, human embryonic stem cells (hESC), human induced pluripotent stem cells (hiPSC), somatic stem cells, cancer stem cells, or any other cell capable of lineage specific differentiation. Specifically described are methods to direct the lineage specific differentiation of hESC and / or hiPSC to nociceptors (i.e. nociceptor cells) using novel culture conditions. The nociceptors made using the methods of the present invention are further contemplated for various uses including, but limited to, use in in vitro drug discovery assays, pain research, and as a therapeutic to reverse disease of, or damage to, the peripheral nervous system (PNS). Further, compositions and methods are provided for producing melanocytes from human pluripotent stem cells for use in disease modeling.

Adipose tissue-derived stromal cells induced to form multicellular aggregates can be formed reliably and consistently, they can be maintained for prolonged periods in adherent or suspension culture, and they are able to survive, grow, and / or differentiate in serum-free media conditions. The present invention provides compositions and methods for the use of such aggregates in tissue repair. This culture platform provides a controlled and defined system in which to study and standardize adult stem cell biology, and begets an instinctive “modular” approach to the predictable replacement and regeneration of adipose tissue.

The invention discloses a method for separating, purifying and culturing human umbilical mesenchymal stem cells and a method for preparing, storing and quickly preparing and applying a wound surface painting reagent of the human umbilical mesenchymal stem cells. The method comprises: firstly, separating, purifying and culturing the human umbilical mesenchymal stem cells; secondly, preparing a methyl cellulose composite culture medium; thirdly, preparing the wound surface painting reagent of the human umbilical mesenchymal stem cells; and fourthly, detecting the biological properties of the human umbilical mesenchymal stem cells. The method takes methyl cellulose as a substrate, mixes the human umbilical mesenchymal stem cells into the methyl cellulose, and generates the wound surface painting reagent of the human umbilical mesenchymal stem cells, and the wound surface painting reagent can be uniformly coated on the wound surface of the skin to form a coating. The invention performs preclinical researches such as animal toxicity test, pyrogen test and hypersensitive test, constructs a cryopreservation resuscitation method for human umbilical mesenchymal stem cells and a stem cell bank, and prepares for large-scale production and preparation of the wound surface painting reagent of the human umbilical mesenchymal stem cells and application of the wound surface painting reagent of the human umbilical mesenchymal stem cells.

The present invention relates to the field of stem cell biology, in particular the lineage specific differentiation of pluripotent or multipotent stem cells, which can include, but is not limited to, human embryonic stem cells (hESC) in addition to nonembryonic human induced pluripotent stem cells (hiPSC), somatic stem cells, stem cells from patients with a disease, or any other cell capable of lineage specific differentiation. Specifically described are methods to direct the lineage specific differentiation of hESC and / or hiPSC into floor plate midbrain progenitor cells and then further into large populations of midbrain fate FOXA2+LMX1A+TH+ dopamine (DA) neurons using novel culture conditions. The midbrain fate FOXA2+LMX1A+TH+ dopamine (DA) neurons made using the methods of the present invention are further contemplated for various uses including, but not limited to, use in in vitro drug discovery assays, neurology research, and as a therapeutic to reverse disease of, or damage to, a lack of dopamine neurons in a patient. Further, compositions and methods are provided for differentiating midbrain fate FOXA2+LMX1A+TH+ dopamine (DA) neurons from human pluripotent stem cells for use in disease modeling, in particular Parkinson's disease. Additionally, authentic DA neurons are enriched for markers, such as CD142, and A9 type neuronal cells.

The invention relates to the field of stem cell biology, specifically to a method for differentiating human pluripotent stem cells into natural killer cells and an application. The invention disclosesa pluripotent stem cell-derived natural killer cell. The pluripotent stem cell-derived natural killer cell expresses CD56, Nkp30, Nkp44 and Nkp46, and also expresses markers CD16 and CD94 of a maturenatural killer cell. The invention also discloses a method for preparing the natural killer cell. The method comprises the following steps: S1, formation of an embryoid body; S2, differentiation of the embryoid body into hematopoietic progenitor cells; S3, differentiation of the hematopoietic progenitor cells into NK cells; and S4, maturation and expansion of the NK cells. With the differentiation method provided by the invention, the pluripotent stem cells can be rapidly, efficiently, simply and conveniently induced to be differentiated into the natural killer cells with low cost on the basis of a culture medium with definite components and an optimized cell factor combination.

The disclosure relates generally to stem cell biology and more specifically to genetic manipulation of stem cells. Methods and compositions using recombinational cloning techniques are disclosed which allow the construction and insertion of complex genetic constructs into embryonic and adult stem cells and progenitor cells. The methods disclosed will allow the harvesting of adult stem cells pre-engineered with integration sites to facilitate early passage genetic modification.

The invention features an “inverse patterning” or “Intaglio-Void / Embed-Relief Topographic (In VERT) molding” manufacturing process for generating high-resolution three-dimensional (3D) multi-cellular microstructures in distinct cellular compartments of a single hydrogel. The platform has general utility in the development of engineered tissues for human therapies, drug testing, and disease models. Additionally, the platform can serve as a model system for studying 3D cell-cell interactions in fields as diverse as stem cell biology to the development of cancer therapeutics.

The present invention relates to the field of stem cell biology, in particular the linage specific differentiation of pluripotent or multipotent stem cells, which can include, but is not limited to, human embryonic stem cells (hESC), human induced pluripotent stem cells (hiPSC), somatic stem cells, cancer stem cells, or any other cell capable of lineage specific differentiation. Specifically described are methods to direct the lineage specific differentiation of hESC and / or hiPSC to nociceptors (i.e. nociceptor cells) using novel culture conditions. The nociceptors made using the methods of the present invention are further contemplated for various uses including, but limited to, use in in vitro drug discovery assays, pain research, and as a therapeutic to reverse disease of, or damage to, the peripheral nervous system (PNS). Further, compositions and methods are provided for producing melanocytes from human pluripotent stem cells for use in disease modeling.

Aspects of the present invention include methods and compositions related to the modulation of molecules regulating the regenerative potential of cells and tissues in the embryonic state and the loss thereof in later fetal and adult stages of development. Said methods and compositions have uses in research in stem cell biology and in increasing regenerative potential in fetal and adult tissues otherwise incapable of regeneration.

The invention relates to the field of stem cell biology, and discloses a preparation method of natural killer cells differentiated from human pluripotent stem cells. The preparation method comprises the steps of S1, forming an embryoid body; S2, performing directional permanent hematopoiesis differentiation on the embryoid body to obtain hematopoietic precursor cells; and S3, differentiating the hematopoietic precursor cells to obtain natural killer cells. The invention also discloses application of the natural killer cell prepared by the method as a pharmaceutical composition for preventing and / or treating tumors by cells. According to the method, efficient and stable differentiation of the NK cells is achieved, EB adherent differentiation is achieved in the induced differentiation period, TGFB inhibitor treatment is prolonged, 10% or above of CD34+CD45+permanent hematopoietic HPCs can be obtained through rapid induction. The method has the advantages of being efficient, stable, low in cost and suitable for large-scale cell preparation production; and the method is easy and convenient to operate, the NK cells obtained through differentiation have typical NK cell surface marking molecules, the anti-tumor function of the NK cells is superior to that of the NK cells derived from umbilical cord blood, and great scientific research and clinical application potentials are achieved.

The invention provides a detection method for the biological efficacy of the umbilical cord mesenchymal stem cell. The method is characterized in that: the immunosuppression ability of the cell is judged by detecting the content of galectin-3 in the umbilical cord mesenchymal stem cell culture supernatant, the pertinence is high, and the quantification is accurate; and moreover, digestive cells are not required, thus the method is suitable for detecting the immunosuppression efficacy in large-scale continuous umbilical cord mesenchymal stem cell culture. In the method provided by the invention, a detection sample is easy to process, and automatic or semi-automatic detection can be performed by use of instrument.

The invention relates to the field of stem cell biology, in particular to midbrain dopaminergic neural precursor cells and a preparation method and application thereof. The midbrain dopaminergic neural precursor cell expresses Lmx1a+, Foxa2+, En1+ and Otx2+, wherein the midbrain dopaminergic neural precursor cell does not express one or more of Nkx2.1, DBX1 and GBX2. The method for preparing the mesencephalic dopaminergic neural precursor cell comprises the following steps: embryoid bodies are formed, midbrain floor cells are obtained, and midbrain dopaminergic neural precursor cells are obtained. By adopting the preparation method, the mesencephalic dopaminergic nerve cells can be rapidly and efficiently differentiated from the human pluripotent stem cells, and the problems are solved that an existing differentiation method is long in time, unstable in effect and low in efficiency, the culture system containing serum or the trophoblast cells are not conducive to the production of subsequent clinical-grade cell preparations and the like.

The invention features an “inverse patterning” or “Intaglio-Void / Embed-Relief Topographic (In VERT) molding” manufacturing process for generating high-resolution three-dimensional (3D) multi-cellular microstructures in distinct cellular compartments of a single hydrogel. The platform has general utility in the development of engineered tissues for human therapies, drug testing, and disease models. Additionally, the platform can serve as a model system for studying 3D cell-cell interactions in fields as diverse as stem cell biology to the development of cancer therapeutics.

The invention provides a detection method for the biological efficacy of the umbilical cord mesenchymal stem cell. The method is characterized in that: the immunosuppression ability of the cell is judged by detecting the content of galectin-3 in the umbilical cord mesenchymal stem cell culture supernatant, the pertinence is high, and the quantification is accurate; and moreover, digestive cells are not required, thus the method is suitable for detecting the immunosuppression efficacy in large-scale continuous umbilical cord mesenchymal stem cell culture. In the method provided by the invention, a detection sample is easy to process, and automatic or semi-automatic detection can be performed by use of instrument.

The invention belongs to the technical fields of stem cell biology and cell culture, and proposes a serum-free culture system for inducing differentiation of multipotent stem cells into neural progenitor cells. The serum-free culture system concretely comprises a serum-free differentiation base culture medium and an additive promoting differentiation of the neural progenitor cells, wherein the serum-free base culture medium comprises the following essential ingredients: a DMEM / F12 culture medium, human serum albumin, L-glutamine, D-glucose, vitamin C, an N2 / B27 cell culture additive and non-essential amino acid; and the additive promoting differentiation of the neural progenitor cells concretely comprises SB431542, dermorphin, fibroblast growth factor-2, retinoic acid and a brain-derived neurotrophic factor. The culture medium provided by the invention can help to increase differentiation of the neural progenitor cells in a cell mixed system and reduce differentiation of the neural progenitor cells into other types of cells, so high-purity neural progenitor cells are finally obtained.

The invention provides a detection method for the biological efficacy of mesenchymal stem cells. The method is characterized by detecting the level of expression secretion type TNFRI in a culture supernatant of the mesenchymal stem cells by an ELISA process to determine the strength of the immunosuppression ability of the cells. The method is easy to operate, high in pertinence and accurate in quantification, and provides a better detection means of the biological efficacy of the mesenchymal stem cells.

The invention provides storage liquid of human umbilical cord mesenchymal stem cells, and belongs to the field of stem cell biology. The storage liquid belongs to a composition of stem cell storage liquid suitable for clinic application. Various active ingredients are used for optimizing the storage conditions, and comprise a mixed water solution containing sodium chloride, albumin, adenosine, angiotensin-II, lithium chloride, flavonoids compounds, heparin, andhuman umbilical cord mesenchymal stem cell sourced exosome, oligopeptide and polypeptide compounds. In the storage liquid, the cell activity of the human umbilical cord mesenchymal stem cells under the condition of 4 DEG C in 3 days is maintained at 90 percent or higher; in addition, the cell phenotype is stable; the cell activity can still be maintained at 90 percent or higher in 7 days under the condition of -20 DEG C; the cell phenotype is also stable. The storage liquid maintains the stable quality of the stem cells; the long-distance conveyance of the stem cells can be realized; the foundation is laid for the wide application of the clinic stem cells.

The invention discloses a stem cell resuscitation device, and belongs to the technical field of stem cell biology. The stem cell resuscitation device comprises a processing table, an adjusting device, a micro shaking device, a placing device and a micro oscillation device, wherein the processing table is arranged on the ground; a water bath kettle is arranged on the processing table, and the adjusting device comprises a rotating part and a lifting part; the rotating part is arranged beside the water bath kettle and arranged on the processing table, and the lifting part is arranged on the rotating part; the micro oscillation device is arranged on the lifting part; the placing device is arranged on the micro oscillation device and right faces the water bath kettle; and the micro oscillation device is arranged in the water bath kettle. According to the stem cell resuscitation device, all the devices are matched with one another for operation, stem cells are heated uniformly, can be resuscitated in batches and can be prevented from being polluted during resuscitation, and the resuscitation effect of the stem cells is improved, the resuscitation quality of the stem cells is improved, and the resuscitation efficiency of the stem cells is improved.

The invention discloses a method for inducing cardiac differentiation of stem cells by utilizing small molecule compounds, relating to the fields of stem cell biology and regenerative medicines. The small molecule compounds refer to a type of carbamate compounds (I) or medicinal salts, hydrates, solvates and isomers thereof. The method comprises the following steps: adding the small molecule compounds into a differential medium to directly act on tested cells, continuously using the small molecule compounds according to the form and growth states of the tested cells and the identification conditions until the expression level of corresponding target genes in the tested cells is increased, and differentiating the tested cells into cardiomyocyte-like cells, wherein the stem cells comprise embryonic stem cells, induced pluripotent stem cells and mesenchymal stem cells. The experiment proves that after the inducers are added into the cell differentiation medium, the myocardial cell differentiation proportion of the stem cells is obviously improved. The differential medium containing a type of carbamate compounds or medicinal salts, hydrates, solvates and isomers thereof can be used for inducing the stem cells to be differentiated into myocardial cells.

This disclosure generally concerns the fields of cell biology and molecular biology. In particular the invention concerns the field of stem cell biology and maturation of stem cell-derived cardiomyocytes. Disclosed is a method for improving the maturity of stem-cell derived cardiomyocytes, in particular of the ventricular type, as can be witnessed by, for example, an improved upstroke velocity of the stem-cell derived cardiomyocytes.

The invention relates to the field of stem cell biology, in particular to a kidney epithelial cell and a preparation method and application thereof. The first disclosed kidney epithelial cell expresses CDH1<+> and PODXL<+>, and at least expresses one of CDH2<+> and LTL<+>. The second disclosed preparation method of the kidney epithelial cell includes the following steps that pluripotent stem cellsare differentiated into posterior streak cells; the posterior streak cells are differentiated into posterior intermediate mesoderms; the posterior intermediate mesoderms are differentiated into kidney progenitor cells; the kidney progenitor cells are differentiated into kidney tubular aggregated precursors; the kidney tubular aggregated precursors are differentiated into kidney vesicles; and thekidney vesicles are differentiated into kidney epithelial cells. The provided preparation method has high differentiation efficiency and stable differentiation effect, and is applied to a variety of hPSC cell lines; and the differentiated nephron progenitor cells can meet the requirements of kidney disease model building, kidney drug toxicity testing, kidney disease related drug screening and other scientific research experiment.

The invention relates to a method for inducing adipose tissue-derived stem cells to differentiate into testicular interstitial-like cells by a single transcription factor, and belongs to the fields of stem cell biology and regenerative medicine. According to the method, chemically modified mRNA of a transcription factor NR5A1 is synthesized in vitro, exosomes extracted from autologous urine of a patient are used as a delivery carrier to form an exosome-modNR5A1 compound, and then the exosome-modNR5A1 compound and ADMSCs derived from autologous tissue of the patient are co-cultured to induce the ADMSCs to be differentiated into Leydig-like cells with a testosterone secretion function. According to the method disclosed by the invention, the Leydig-like cells with the testosterone secretion function can be quickly and efficiently obtained, meanwhile, the cells can effectively react to the stimulation of luteinizing hormone, and the cells have a gene expression profile related to the androgen synthesis of mature Leydig cells. The Leydig cell obtained by the invention has a good application prospect in the aspect of treatment of male hypogonadism.

The invention belongs to the technical field of stem cell biology and cell culture and provides a serum-free culture system capable of promoting induced pluripotent stem cells to be differentiated tocortical nerve cells. The serum-free culture system specifically comprises a serum-free differentiation basic culture medium and additives capable of promoting cortical nerve cell differentiation, andnecessary ingredients of the serum-free basic culture medium include DMEM / F12 culture medium, human serum albumin, L-glutamine, D-glucose, vitamin C, N2 / B27 cell culture additive and non-essential amino acid. The additives capable of promoting cortical nerve cell differentiation specifically include SB431542, dermorphin, fibroblast growth factor-2, retinoic acid, BDNF, GDNF, CTNF, DAPT and cytarabine. By using the culture medium, cortical nerve cell differentiation in a cell mixed system can be promoted, and differentiation towards cells of other types is reduced, so that high-purity corticalnerve cells are obtained finally.

The invention discloses a method for separating, purifying and culturing human umbilical mesenchymal stem cells and a method for preparing, storing and quickly preparing and applying a wound surface painting reagent of the human umbilical mesenchymal stem cells. The method comprises: firstly, separating, purifying and culturing the human umbilical mesenchymal stem cells; secondly, preparing a methyl cellulose composite culture medium; thirdly, preparing the wound surface painting reagent of the human umbilical mesenchymal stem cells; and fourthly, detecting the biological properties of the human umbilical mesenchymal stem cells. The method takes methyl cellulose as a substrate, mixes the human umbilical mesenchymal stem cells into the methyl cellulose, and generates the wound surface painting reagent of the human umbilical mesenchymal stem cells, and the wound surface painting reagent can be uniformly coated on the wound surface of the skin to form a coating. The invention performs preclinical researches such as animal toxicity test, pyrogen test and hypersensitive test, constructs a cryopreservation resuscitation method for human umbilical mesenchymal stem cells and a stem cell bank, and prepares for large-scale production and preparation of the wound surface painting reagent of the human umbilical mesenchymal stem cells and application of the wound surface painting reagent of the human umbilical mesenchymal stem cells.

The invention relates to the technical field of stem cell biology, in particular to a detection method for in-vitro Th1 and Th17 inhibition of mesenchymal stem cells. The method comprises the following steps: 1, co-culturing mesenchymal stem cells and peripheral blood mononuclear cells; 2, carrying out fixed membrane rupture dyeing and detection. According to the detection method provided by the invention, lymphocyte proliferation and differentiation are promoted through co-culture of mesenchymal stem cells and peripheral mononuclear cells, so that the expression of the inhibition effect of the mesenchymal stem cells is promoted, and the inhibition effect is improved. According to the method, the PBMC cells obtained through co-culture are stimulated and blocked, so that factor secretion is promoted, meanwhile, inhibition and factor blocking are added for detection, so that expression of Th1 and Th17 in the PBMC cells is further promoted, expression of Th1 and Th17 obtained through detection is improved, and the inhibition effect is further improved.

The invention belongs to the technical fields of stem cell biology and cell culturing, and provides a serum-free culturing system inducing differentiation of pluripotent stem cells into motor neuron cells. The serum-free culturing system specially comprises a serum-free differentiation basic culture medium and an additive for promoting the differentiation of the motor neuron cells, wherein necessary components of the serum-free differentiation basic culture medium comprise a DMEM / F12 culture medium, a human serum albumin, L-glutamine, D-glucose, vitamin C, N2 / B27 cell culture additive and unnecessary amino acid. The additive for promoting the differentiation of the motor neuron cells specially comprises SB431542, dermomorphin, fibroblast growth factors-2, retinoic acid, BDNF, GDNF, CTNF, DAPT, Smoothened agonist, and cytarabine. The culture medium is adopted to help improve the differentiation of the motor neuron cells in a cell mixing system, the differentiation of other types of cells is reduced, and finally, the motor neuron cells with high purity can be obtained.

The present invention relates to a composition for increasing the biological activity of stem cells using mixture 4F. Stem cells treated with mixture 4F according to the present invention not only acquire stemness, but also have the effect of improving cell proliferative ability and mobility, and thus, after being transplanted into a body, the stem cells can improve cell survival and engraftment and further enhance the ability to regenerate blood vessels and tissues. Accordingly, the stem cells can have various applications in the fields of stem cell differentiation and ischemic disease prevention or treatment.

The invention relates to the field of stem cell biology, in particular to a method and application for differentiating human pluripotent stem cells into natural killer cells. The invention discloses a natural killer cell derived from pluripotent stem cells, which expresses CD56, Nkp30, Nkp44 and Nkp46, and also expresses markers CD16 and CD94 of mature natural killer cells. The invention also discloses a method for preparing natural killer cells, comprising the following steps: S1: forming embryoid bodies; S2: differentiation of embryoid bodies to hematopoietic progenitor cells; S3: differentiation of hematopoietic progenitor cells to NK cells; S4: NK cells maturation and expansion. The differentiation method provided by the present invention is based on a culture medium with defined components and an optimized combination of cytokines, which can rapidly, efficiently, simply and cost-effectively differentiate induced pluripotent stem cells into natural killer cells.

The present invention relates to the field of stem cell biology, in particular the lineage specific differentiation of pluripotent or multipotent stem cells, which can include, but is not limited to, human embryonic stem cells (hESC) in addition to nonembryonic human induced pluripotent stem cells (hiPSC), somatic stem cells, stem cells from patients with a disease, or any other cell capable of lineage specific differentiation. Specifically described are methods to direct the lineage specific differentiation of hESC and / or hiPSC into floor plate midbrain progenitor cells and then further into large populations of midbrain fate FOXA2+LMX1A+TH+ dopamine (DA) neurons using novel culture conditions. The midbrain fate FOXA2+LMX1A+TH+ dopamine (DA) neurons made using the methods of the present invention are further contemplated for various uses including, but not limited to, use in in vitro drug discovery assays, neurology research, and as a therapeutic to reverse disease of, or damage to, a lack of dopamine neurons in a patient. Further, compositions and methods are provided for differentiating midbrain fate FOXA2+LMX1A+TH+ dopamine (DA) neurons from human pluripotent stem cells for use in disease modeling, in particular Parkinson's disease. Additionally, authentic DA neurons are enriched for markers, such as CD142, and A9 type neuronal cells.

The present invention relates to the field of stem cell biology, in particular the linage specific differentiation of pluripotent or multipotent stem cells. Specifically described are methods to direct the lineage specific differentiation of hiPSC to sensitive neurons or neuronal fibers innervating the human skin, such as neural crest stem cells (NCPCs) and here called peripheral sensory neurons (PSNs) using novel culture conditions. It is also described a method for screening a biological agent in vitro. The PSNs obtained using the methods of the present invention are further contemplated for various uses including, but limited to, use in in vitro tests or disease modelling, such as drug discovery assays, cell therapy on a higher scale, detecting a range of skin irritants and other compounds of interest, for studying skin aging mechanism, and for producing a dermocosmetic product. Also, it is contemplated the use of ligands in vitro for checking the differentiation and / or activation of the PSN cells, produced by the method of the invention, and the PSN cells, produced by the method of the invention.