152 results about "Directed differentiation" patented technology

The invention belongs to the field of stem cell biology, and relates to pedigree specific differentiation of human pluripotent or embryonic stem cells, in particular to a mesenchymal stem cell induced differentiation culture medium and application thereof. The mesenchymal stem cell induced differentiation culture medium is used for directly inducing and differentiating human induced pluripotent stem cells / or embryonic stem cells into mesenchymal stem cells, the culture medium comprises a mesenchymal stem cell basic culture medium and an additive, and the additive comprises the following components: 2-10 [mu] M of a TGF-beta inhibitor, 2-10 [mu] M of a GSK3 beta inhibitor and 5-10 [mu] M of a Rock inhibitor. By adopting the induced differentiation culture medium provided by the invention, human induced pluripotent stem cells / or embryonic stem cells can be directly induced and differentiated into mesenchymal stem cells, and the obtained mesenchymal stem cells have the advantages of infinite sources, stable quality, high homogeneity and the like; and the human induced pluripotent stem cells / or embryonic stem cells can be directly and directionally differentiated into the mesenchymal stem cells in a relatively short time, so that the culture time is shortened, and the production cost is reduced.

Disclosed in the present invention is a use of alisol-B23-acetate (AB23a) in the preparation of a drug for preventing or treating hypertrophic cardiomyopathy and diseases caused by the hypertrophic cardiomyopathy. The present invention provides for the first time a use of AB23a in the prevention or treatment of hypertrophic cardiomyopathy. According to the present invention, use of AB23a in direct in vitro treatment of cardiomyocytes derived from directed differentiation of human embryonic stem cells revealed that AB23a-based in-vitro treatment can significantly inhibit hypertrophic phenotypes of the human embryonic stem cells-cardiomyocytes; using an AB23a-containing feed to feed mice with hereditary hypertrophic cardiomyopathy caused by a gene mutation revealed that AB23a prominently relieves pathological myocardial hypertrophy of the mice and improves the cardiac function; AB23a can be used for preparing a drug for resisting hereditary hypertrophic cardiomyopathy, offering a novel way and means for treating hypertrophic cardiomyopathy; and AB23a is the main medicinal ingredient in the traditional Chinese medicinal herb-Rhizoma Alismatis, is safe to organisms, and has good clinical application prospects.

The invention discloses a method for differentiating induced pluripotent stem cells into megakaryocytes and platelets, and relates to the technical field of induced pluripotent stem cell directional differentiation, and the method comprises the following steps: inoculating an iPSC single cell into a culture vessel, and carrying out adherent culture through a culture solution; after the cell adheres to the wall and before cloning is not formed, the liquid is changed into a first induction culture liquid, and the cell is induced and differentiated into a mesoderm cell; changing the solution into a second induction culture solution to induce and differentiate the mesoderm cells into hematopoietic endothelial cells; collecting the suspension cells, culturing the suspension cells in an amplification culture solution, and enriching megakaryocytes; inducing megakaryocyte polyploidy through a third induction culture solution; and culturing the polyploidy megakaryocyte through a fourth induction culture solution to promote platelet release. The method for inducing the pluripotent stem cells to differentiate into megakaryocytes and platelets is convenient and rapid, the megakaryocytes and platelets can be obtained through iPSC in a short time, and feeder cells, serum and additional gene editing are not needed; meanwhile, the yield is high.

The invention discloses a biconvex crystalline lens organ relying on stiffness-controllable hydrogel external directional induction as well as a construction method and application thereof. A crystalline lens organ model provided by the invention is jointly realized by virtue of stiffness-controllable hydrogel and a composite growth factor; and the pluripotent stem cells are rapidly and directionally induced into crystalline lens organs with high quality in vitro. According to the technical scheme, the method comprises the following steps: (1) directionally inducing and differentiating the pluripotent stem cells into primary ectoderm cell clusters; (2) preparing hydrogel with specific rigidity and carrying out surface cell adhesion treatment; (3) planting the primary ectoderm cells on the cleaned polyacrylamide hydrogel and inducing the primary ectoderm cells into a neural ectoderm cell cluster; and (4) inducing the neural ectoderm cells to directionally differentiate into mature crystalline lens organs. The three-dimensional lens organ-like model obtained by the method has a sphere-like biconvex morphology with a top surface and a bottom surface, is transparent and has structures such as capsule membranes, lens epithelial cells and lens fibrocytes, and the bottom surface is also wrapped by a complete lens capsule membrane. Compared with the prior art, the method realizes the formation of a complete capsule film on the bottom surface and a sphere-like form. The obtained crystalline lens organ can be used for research on crystalline lens embryonic development, research on cataract pathogenesis and research, development and screening of cataract drugs.

The invention relates to a composite hydrogel artificial bile duct as well as a preparation method and application thereof, the artificial bile duct is prepared by taking sodium alginate (SA) loaded with human umbilical cord mesenchymal stem cells and methylacryloylated gelatin (GelMA) composite hydrogel as raw materials, and the artificial bile duct can be used for biological materials or instruments for treating bile duct defects. The composite hydrogel artificial bile duct has better biocompatibility, can be tightly connected with an in-vivo tissue mucous membrane, and reduces stimulation to a local mucous membrane, so that bile duct obstruction caused by local tissue granulation hyperplasia is avoided, and adverse reactions are reduced; in addition, the composite hydrogel artificial bile duct can be automatically degraded after being implanted into a human body; the structure is compact, the mechanical property is good, and a good environment is provided for cell proliferation; the human umbilical cord mesenchymal stem cells can be directionally differentiated into bile duct cell-like cells, and bile duct injury repair is promoted.

The invention provides application of enhanced mesenchymal stem cells and exosomes in chronic nephritis and renal failure, and belongs to the technical field of biological medicine manufacturing. Through composite pretreatment of salidroside and astragalus polysaccharide, the treatment potential of exosomes secreted by the mesenchymal stem cells is remarkably enhanced, a new thought is provided for intervention of the chronic kidney diseases, and various pathologies of the chronic kidney diseases are covered. In addition, a 3D microenvironment constructed by fibrin gel is combined to simulate an in-vivo extracellular matrix, the natural growth state of the stem cells is better fitted, directional differentiation and function regulation of the stem cells are promoted, and therefore the biological activity and targeting performance of the exosome are improved.

The present invention relates to an induced pluripotent stem cell (iPSC) or a somatic cell derived from an iPSC by forward programming or directed differentiation constitutively expressing an alternative splice form of Lamin A. The cells comprise an expression cassette comprising a coding sequence for the alternative Lamin A splice form operably linked to a cumate operator (CuO) sequence and a constitutive promoter. Also provided is the use of the cells described herein in a method of identifying a gene or combination of genes involved in the reversal of an ageing phenotype or in the maintenance of a non-aged phenotype. Methods of rejuvenating somatic cells, treating diseases or disorders associated with ageing, and modulating ageing (e.g. age-modulating methods and uses), including in vivo, are also provided, said methods comprising modulating the expression and / or activity of the gene / combination of genes identified herein.

The invention relates to the technical field of cell differentiation, in particular to a method for directionally differentiating iPSC into iNK. Specifically, the method for preparing the CD34 + hematopoietic stem / progenitor cell and the iNK on the shaking table comprises the following steps: forming an embryoid (EB) by the iPSC on the shaking table, and then collecting a single cell supernatant to obtain the iNK. According to the method, high-proportion CD34 < + > hematopoietic stem / progenitor cells are obtained by adjusting the factor concentration and collecting the supernatant and are further induced and differentiated into the iNK, and more than 100 iNKs can be harvested before amplification of one iPSC according to the iNK harvested in batches.

According to the application of the dipsacus asper saponin VI in regulating and controlling the differentiation of the surface stem cells, the brand new pharmacological action of the dipsacus asper saponin VI in targeted promotion of the differentiation of the epidermal stem cells is found for the first time, the cognitive limitation of the prior art is broken through, a brand new target spot and direction are provided for research and development of innovative drugs for wound repair, and the application of the dipsacus asper saponin VI in regulating and controlling the differentiation of the epidermal stem cells is realized. A medicine containing teasel saponin VI is adopted, epidermal stem cells are taken as a core target, wound reepithelialization is accelerated by promoting directional differentiation of the epidermal stem cells, the mechanism is clear, the targeting property is high, a core link capable of directly hitting wound healing is adopted, epidermal stem cell differentiation is regulated in a targeted manner, and meanwhile, the medicine has the synergistic effects of resisting inflammation and promoting angiogenesis; the obvious application potential is realized on refractory wounds such as diabetic feet and pressure sores.

The invention relates to a cellulose-based hydrogel prepared based on a cellulose copolymer, a cellulose derivative copolymer or a blend thereof, ultramicro fibers of cellulose form a net structure, and the cellulose-based hydrogel has a highly-ordered three-dimensional micro-nano structure, is better in transparency and is more beneficial to observation of a cell state. The cellulose-based hydrogel is suitable for culture of 3D cell culture tissues and organoids, the stem cells cultured by the cellulose-based hydrogel can form cluster culture stem cells and organoids, the operation is simple, compact fused monolayer endoderm cells are formed, non-directional differentiation of the stem cells can be inhibited, and the formed organoids are thick in vesicle wall and are in a solid shape. And the obtained product is a differentiated mature organ and can be used for high-throughput drug screening, drug toxicity testing and regenerative medicine.

The generation of complex organ tissues from human embryonic and pluripotent stem cells (PSCs) remains a major challenge for translational studies. It is shown that PSCs can be directed to differentiate into intestinal tissue in vitro by modulating the combinatorial activities of several signaling pathways in a step-wise fashion, effectively recapitulating in vivo fetal intestinal development. The resulting intestinal “organoids” were three-dimensional structures consisting of a polarized, columnar epithelium surrounded by mesenchyme that included a smooth muscle-like layer. The epithelium was patterned into crypt-like SOX9-positive proliferative zones and villus-like structures with all of the major functional cell types of the intestine. The culture system is used to demonstrate that expression of NEUROG3, a pro-endocrine transcription factor mutated in enteric anendocrinosis is sufficient to promote differentiation towards the enteroendocrine cell lineage. In conclusion, PSC-derived human intestinal tissue should allow for unprecedented studies of human intestinal development, homeostasis and disease.

Provided are methods and compositions for obtaining functionally enhanced derivative effector cells obtained from directed differentiation of genomically engineered iPSCs. The derivative cells provided herein have stable and functional genome editing that delivers improved or enhanced therapeutic effects. Also provided are therapeutic compositions and uses thereof comprising the functionally enhanced derivative effector cells alone, or with antibodies or checkpoint inhibitors in combination therapies.

This invention discloses the application of alismazone B acetate in the preparation of drugs for the prevention or treatment of hypertrophic cardiomyopathy and its related symptoms. This invention is the first to propose the use of alismazone B acetate (AB23a) for the prevention or treatment of hypertrophic cardiomyopathy. The invention utilizes AB23a to directly treat cardiomyocytes obtained from directed differentiation of human embryonic stem cells in vitro, finding that AB23a treatment significantly inhibits the hypertrophic phenotype of human embryonic stem cell-cardiomyocytes. Feeding mice with hereditary hypertrophic cardiomyopathy caused by gene mutations to a diet containing AB23a showed that AB23a significantly alleviated pathological myocardial hypertrophy and improved cardiac function in mice. AB23a can be used to prepare drugs against hereditary hypertrophic cardiomyopathy, providing a new approach and method for treating hypertrophic cardiomyopathy. AB23a is the most important medicinal component of the traditional Chinese medicine Alisma plantago-aquatica, is safe for organisms, and has good clinical application prospects.

The present invention belongs to the technical field of cell biology, and particularly relates to the application of Hippo-Yap pathway regulators in promoting the directed differentiation of induced pluripotent stem cells into pancreatic progenitor cells. The method of the present invention can efficiently obtain pancreatic progenitor cells derived from non-human primate rhesus monkeys in vitro, providing a material source for further obtaining functional pancreatic islet β cells of non-human primate rhesus monkeys, and more importantly, providing a practical basis for the autologous transplantation treatment of non-human primate diabetic animal models.

The invention discloses a system for directionally differentiating chicken primordial germ cells into gametes in vitro and a regulation and control method, the system comprises a basic culture medium: based on DMEM / F12, adding 10% of fetal calf serum, 2% of chicken serum, 2 mmol / L of L-glutamine, 1 mmol / L of sodium pyruvate and 5.5 * 10 <-5 > mol / L of beta-mercaptoethanol; the spermatogonium induction culture medium further comprises BMP4, RA and GDNF; the oogonium induction culture medium also comprises BMP15, GDF9 and EGF (Epidermal Growth Factor). According to the system and the regulation and control method, the differentiation efficiency is improved, the culture period is shortened, and the number of cell clusters is remarkably increased.

The invention disclosed herein generally relates to methods and systems for converting stem cells into specific tissue(s) or organ(s) through directed differentiation. In particular, the invention disclosed herein relates to methods and systems for promoting vascularized human intestinal organoid tissue having an intestine-specific EC transcriptional signature from hindgut spheroid tissue produced in vitro from the described methods.

The invention discloses a female vaginal mucosa cell maintenance preparation and a preparation method and application thereof, and belongs to the technical field of biological agents, the female vaginal mucosa cell maintenance preparation is prepared from dental pulp mesenchymal stem cells, portulaca oleracea polysaccharide, lactic acid, quercitrin, glycerin and stem cell carriers, and vaginal injection paste is obtained by evenly mixing the dental pulp mesenchymal stem cells, the portulaca oleracea polysaccharide, the lactic acid, the quercitrin, the glycerin and the stem cell carriers through a proper amount of deionized water. The purslane polysaccharide and the dental pulp mesenchymal stem cells form a unique synergistic effect, through physical wrapping, the survival rate and delivery efficiency of the stem cells in a vaginal cavity are improved, local inflammatory response is inhibited, a microenvironment is optimized to support directional differentiation of the stem cells into epithelial cells, paracrine activity of the stem cells is enhanced, and repairable factors are released to promote cell-level regeneration. The problem of hormone dependence is solved, deep maintenance of targeted mucous membrane cells is achieved, limitation of an existing therapy is broken through by directly nourishing cells, activating and regenerating, enhancing barrier integrity and maintaining an acidic microenvironment, and an ideal treatment effect is achieved.

The invention relates to hair-blacking essence and a preparation method thereof. Screening a symbiotic bacterium metabolism extract with small molecule peptide metabolism regulation capability; by combining low-concentration azelaic acid and zinc lactate components, scalp grease oxidation and abnormal proliferation of propionibacterium are inhibited; the expression of IL-10 anti-inflammatory factors is induced, the expression of TNF-alpha and IL-6 is reduced, and the hair follicle microinflammation state is reduced; a Wnt / beta-catenin pathway activator is used for inducing the melanin stem cells to be in a static state; the FGF-2 and the EGF are combined in a micro-dose manner to promote the amplification into precursor cells; mITF regulation peptides and a small amount of trace copper ions are supplemented to induce directional differentiation into mature melanocytes; a microcapsule carrier system with a three-layer structure; the penetration efficiency is improved; antioxidative peptide rich in reduced glutathione, NAD and a promoting factor are used, so that melanocytes are protected from oxidative damage; a coenzyme Q10 type mitochondrial homeostasis regulator is introduced, so that the energy metabolism is improved, the cell aging is delayed, the targeting property is strong, and the effect is accurate.

The invention discloses an analysis method for storage of single vesicles in ventral spinal cord motor neuron-like organs induced and differentiated by an iPSC technology. The method sequentially comprises the steps of construction of an acetylcholine nano microelectrode sensor interface, recovery and culture of human pluripotent stem cells, directional differentiation of spinal cord movement organs and storage and detection of single vesicle acetylcholine. A new thought and a new method are provided for monitoring the level of acetylcholine stored in the single vesicles at the single cell level, and the method has important theoretical significance and wide practical value in research of pathogenesis of diseases, drug screening and the like in the field of neuroscience.

The invention relates to the technical field of cell differentiation, in particular to an ovarian granular cell exosome preparation method and system based on cell directional differentiation, and the method comprises the following steps: reprogramming starting material cells to obtain initial induced pluripotent stem cells, carrying out cell identification to obtain induced pluripotent stem cells, and carrying out directional differentiation on the induced pluripotent stem cells to obtain the ovarian granular cell exosome. Obtaining an ovarian granular cell cluster, obtaining a culture supernatant, purifying the culture supernatant to obtain an initial exosome, carrying out quality detection on the initial exosome to obtain a detection result, if the detection result is that the detection does not reach the standard, obtaining an adjusted exosome, taking the adjusted exosome as the initial exosome, and taking the adjusted exosome as the initial exosome. And returning to the step of detecting the quality of the initial exosome until the detection result is that the detection reaches the standard, and confirming the initial exosome as the target exosome when the detection result is that the detection reaches the standard. The problem that the purity of the exosome is low in the preparation process of the ovarian granular cell exosome at present can be solved.

The invention provides a method for producing functional spinal cord GABAergic neural precursor cells from human pluripotent stem cells, which successfully realizes directional differentiation of the human pluripotent stem cells by adopting a feeder-layer-free culture method based on a three-stage in-vitro induced differentiation strategy. The invention further provides application of the cells obtained through the method in treatment of spinal cord injury, a novel strategy based on the functional spinal cord GABAergic neural precursor cells is provided, a new thought and means are provided for treatment of spinal cord injury, and stem cell treatment is expected to proceed towards clinical precision and industrial application.

The invention discloses a method for inducing iPSCs (induced pluripotent stem cells) to directionally differentiate into vascular endothelial cells by using a small molecule compound and cell factor composition, which is characterized in that iPSCs cells are used as starting cells, and a vascular endothelial cell differentiation scheme completely dependent on chemical small molecule and cell factor stimulation is established. An E6 culture medium containing CHIR-99021, BMP4 and Activin A is utilized to induce mesoderm progenitor cells, an E6 culture medium containing VEGFA (vascular endothelial growth factor A), FGF-2, 8-Bromoadenosine 3 ', 5'-cyclic monophosphate monophosphate, SB431542, Forskolin and Melatonin is utilized to induce the endothelial cells, and an E6 culture medium containing VEGFA, FGF-2 and Hydrocortisone is utilized to further induce and differentiate the endothelial cells into the vascular endothelial cells. By using the scheme, the differentiation efficiency can exceed 99%, and differentiated cells have phenotypes and functions of vascular endothelial cells and do not express pluripotent related markers SSEA-4, TRA-1-60 and TRA-1-81.

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.

The invention relates to the technical field of biomedicine, and discloses an application of a light-driven organic semiconductor nano motor in preparation of a product for promoting differentiation of neural stem cells, and the organic semiconductor nano motor is obtained by carrying out surface treatment on poly (3-hexylthiophene) through lauryl sodium sulfate and normal propyl alcohol. In-vivo experiments find that in-situ injection of the P3HT nanomotor in a subventricular region (V-SVZ) of a mouse can maintain stable photoresponse performance under visible light irradiation, and can promote directional differentiation of neural stem cells to a neural pedigree (rather than glial cells). In addition, the organic semiconductor nano motor has good cytocompatibility and systemic biological safety, and has a good application prospect in the field of nerve regeneration medicine.

The invention discloses an enzyme response magnetic resonance imaging nanoprobe as well as a preparation method and application thereof. The enzyme response magnetic resonance imaging nanoprobe comprises a metal nanoparticle with a surface functional group A as an imaging basic unit, and a polypeptide sequence with functional groups B at two ends as a cross-linking agent and an enzyme response unit, the enzyme response magnetic resonance imaging nano-probe is a metal nano-particle cross-linked body formed by cross-linking the metal nano-particles through a polypeptide sequence. The enzyme response magnetic resonance imaging nanoprobe disclosed by the invention can be specifically cut by a high-expression enzyme in a cell differentiation process to form monodispersed metal nanoparticles, so that signal transformation is presented on magnetic resonance imaging; meanwhile, the enzyme response magnetic resonance imaging nanoprobe can establish effective association among the probe existence form, the cell state and the imaging signal, and is used for tracking the directional differentiation process of mesenchymal stem cells, neural stem cells and promyelocytic leukemia cells in real time.

The invention relates to a gradient scaffold material based on particle gel as well as a preparation method and application thereof. The preparation method comprises the following steps: (1) constructing a shaping framework; (2) respectively preparing a plurality of hydrogel particles with different properties; and (3) respectively injecting the hydrogel particles with different properties into the shaping frame, and carrying out in-situ crosslinking on the hydrogel particles to prepare the gradient scaffold material. The scaffold material prepared by the method has the characteristic that the composition, the structure and the mechanical property are continuously adjustable in space, and the scaffold also has good biocompatibility and a penetrating porous structure, so that inward migration, proliferation and directional differentiation of cells are facilitated, and the scaffold material has good application prospects. The method has important application in the fields of bone-cartilage, tendon-bone, skin and other gradual change tissue engineering interface repair and the like.

Provided is a method for differentiating pluripotent stem cells in vitro to prepare a cell population mainly composed of midbrain floor plate dopaminergic progenitor cells, wherein the progenitor cells can efficiently and directionally differentiate into midbrain dopaminergic neuron precursors and midbrain dopaminergic neurons after being transplanted into a subject. Further provided is a cell population prepared by the method, a pharmaceutical composition containing the cell population, and a method for using the cell population or the pharmaceutical composition to treat or ameliorate a neurological disorder caused by a reduction in the number of midbrain dopaminergic neurons and / or impairment of function thereof.

The invention provides an application and a method of GSH in promoting mature differentiation of hiPSC-CMs, and the method is characterized in that in the process of directionally differentiating the hiPSC-CMs into the hiPSC-CMs, an effective dose of GSH is added into a differential medium, and the optimal concentration is 20-100 [mu] M. According to the invention, through simple and economical exogenous supplement of GSH, maturation of hiPSC-CMs is systematically promoted from multiple dimensions, including enhancement of intracellular anti-oxidation defense capability; energy metabolism is promoted to be converted from glycolysis to oxidative phosphorylation; up-regulating expression of cardiac muscle cell specific structural protein and calcium ion channel related genes; mitochondrial biological generation and functions are promoted; and the cell morphological structure is induced to develop to a mature phenotype. The method is simple and convenient to operate and high in biocompatibility, the hiPSC-CMs with mature functions can be stably obtained, and a high-quality cell source is provided for application of the hiPSC-CMs in the fields of heart disease modeling, drug cardiotoxicity screening and the like.

The invention discloses a method for inducing directional differentiation of stem cells by neural factors as well as a preparation and application thereof, and the preparation is a liquid preparation and comprises the following components: BDNF with the concentration of 25-75 ng / mL, GDNF with the concentration of 20-30 ng / mL, NT-3 with the concentration of 5-10 ng / mL and TSA with the concentration of 100-150 nM. The preparation for inducing the directional differentiation of the stem cells through the neural factors can remarkably improve the directional differentiation efficiency of the neural stem cells to neurons, the purity of the neurons is at a high level, the cell viability is good, target cells with mature functions can be efficiently induced, and a new thought is provided for cell treatment of Parkinson's disease.

The application discloses an enzyme-responsive magnetic resonance imaging nano probe and a preparation method and application thereof. The enzyme-responsive magnetic resonance imaging nano probe comprises a surface functional group A metal nanoparticle as an imaging basic unit, and a functional group B polypeptide sequence as a crosslinking agent and an enzyme-responsive unit; and the enzyme-responsive magnetic resonance imaging nano probe is a metal nanoparticle crosslinking body formed by crosslinking the metal nanoparticle through the polypeptide sequence. The enzyme-responsive magnetic resonance imaging nano probe can be specifically cut by an enzyme highly expressed in a cell differentiation process, and forms a monodispersed metal nanoparticle, so that a signal change is presented on magnetic resonance imaging. Meanwhile, the enzyme-responsive magnetic resonance imaging nano probe can establish an effective correlation among a probe existing form, a cell state and an imaging signal, and is used for real-time tracking of a directional differentiation process of mesenchymal stem cells, neural stem cells and promyelocytic leukemia cells.