81 results about "Directed differentiation" patented technology

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.

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 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.

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 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 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.

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.

This invention discloses the application of prostaglandin PGE1 or PGE2 in the preparation of products that promote the proliferation of exhausted precursor T cells. Specifically, by adding prostaglandin PGE1 or PGE2 to a T cell culture system and culturing, highly efficient expansion of exhausted precursor T cells can be obtained, while simultaneously inhibiting CD8. + The degree of T cell exhaustion is such that when prostaglandin PGE1 or PGE2 is discontinued, exhausted precursor T cells can rapidly differentiate into CD8 cells. + T cells. Therefore, using prostaglandin PGE1 or PGE2 in the in vitro preparation process of adoptive T cell therapy can significantly increase CD8+. + Increasing the total number of T cells, improving the proportion of Tpex cells, and reducing the expression of exhaustion markers provide technical support for the clinical promotion of tumor-infiltrating lymphocyte therapy for solid tumors.

The invention relates to the technical field of regenerative medicine application, in particular to a serum-free induction culture medium with definite components for promoting adipose tissue-derived cells to be differentiated into dermal papilla cells, an induction method and application. The invention relates to a method for promoting adipose tissue-derived cells to differentiate into dermal papilla cells, which is characterized in that the adipose tissue-derived cells are induced and cultured by using a serum-free induction culture medium, so that the adipose tissue-derived cells are differentiated into the dermal papilla cells. The formula of the serum-free induction culture medium comprises hepatocyte growth factors, hydrocortisone, bone morphogenetic protein 2, BIO and basic fibroblast growth factors. Based on the differentiation potential of adipose tissue-derived mesenchymal cells and the function maintenance mechanism of dermal papilla cells, a serum-free induction system with definite components is constructed, and directional differentiation is realized. According to the technical scheme, the technical problem that a method for inducing the adipose-derived mesenchymal cells to be directionally differentiated into the dermal papilla cells with the effective hair follicle regulation function is lacked in the prior art can be solved, and the method has ideal application and popularization potential.

Use of a TGF-β small molecule inhibitor in the field of nerve regeneration, for in vitro regeneration and directed differentiation of a variety of nerve cells and brain-like organs. By adding the TGF-β small molecule inhibitor to a set of basal media having clear chemical compositions, pluripotent stem cells can be induced into adult cells derived from a variety of neural stem cells, and the number of induced nerve cells can be greatly increased.

Provided are cortical interneurons and other neuronal cells and in vitro methods for producing such cortical interneurons and other neuronal cells by the directed differentiation of stem cells and neuronal progenitor cells. The present disclosure relates to novel methods of in vitro differentiation of stem cells and neural progenitor cells to produce several type neuronal cells and their precursor cells, including cortical interneurons, hypothalamic neurons and pre-optic cholinergic neurons. The present disclosure describes the derivation of these cells via inhibiting SMAD and Wnt signaling pathways and activating SHH signaling pathway. The present disclosure relates to the novel discovery that the timing and duration of SHH activation can be harnessed to direct controlled differentiation of neural progenitor cells into either cortical interneurons, hypothalamic neurons or pre-optic cholinergic neurons. The present disclosure also relates to compositions of cortical interneurons, hypothalamic neurons or pre-optic cholinergic neurons, and their precursors, that are highly enriched and can be used in variety of application. These cells can be used therapeutically to treat neurodegenerative and neuropsychiatric disorders, and can be used for disease modeling and drug screening.

This method discloses a technique for inducing the directed differentiation of human amniotic mesenchymal stem cells into liver organoids, comprising the following steps: S1, expanding and culturing third-generation human amniotic mesenchymal stem cells; S2, discarding the culture supernatant from step S1 and adding a first induction medium; S3, discarding the culture supernatant from step S2 and adding a second induction medium; S4, discarding the culture supernatant from step S3, adding trypsin to digest the cells, resuspending the cells in a third induction medium, centrifuging, and culturing; S5, after the completion of step S4, discarding the culture supernatant and adding a fourth induction medium, ultimately obtaining functionally mature liver organoids. This induction method avoids the tumorigenicity problems associated with other methods, such as induced pluripotent stem cell induction, making it more suitable for industrialization and clinical application. Furthermore, it uses simple components and has lower technical requirements, thus greatly increasing the feasibility of the experiment and facilitating industrialization.

This invention discloses a method for the isolation, culture, and identification of chicken sexual gonadal cells, belonging to the field of biotechnology. The invention selects chicken embryos hatched to 18.5 days old, isolates the gonads after PCR sex identification, and uses 0.25% trypsin-EDTA digestion followed by filtration through a 70μm filter to obtain a single-cell suspension with a survival rate >90% and free from PGC contamination. A sex-specific culture system is established, allowing cells to be stably passaged and maintain a sex-specific phenotype. Through morphological, qRT-PCR, immunofluorescence, and Western blot multidimensional identification, accurate identification of male testicular Sertoli cells and female ovarian granulosa cells is achieved. This method is simple to operate, has good reproducibility, and produces high cell viability and purity. It can provide a stable gonadal microenvironment for the in vitro directed differentiation of chicken primordial germ cells, significantly improving gamete induction efficiency. It is suitable for research on avian reproductive and developmental mechanisms, transgenic breeding, and germplasm resource preservation.

The invention relates to the technical field of biomedicine, in particular to a method for preparing an acellular matrix from novel oligopeptide MY-1 and application of the acellular matrix. The preparation method comprises the following steps: (1) separation and culture of mesenchymal stem cells; (2) culturing under the condition of an MY-1 stimulating factor to obtain an extracellular matrix; and (3) carrying out decellularization treatment to obtain the decellularized matrix. The preparation method is simple, the prepared specific dECM can remarkably promote in-vitro migration and directional differentiation of mesenchymal stem cells, osteoporotic femoral defect repair can be highly promoted, and the performance of the dECM is better than that of dECM cultured through conventional culture or addition of PTH or knock-down type III collagen.

The invention discloses application of prostaglandin PGE1 or PGE2 in preparation of a product for promoting proliferation of depleted precursor T cells. Specifically, prostaglandin PGE1 or PGE2 is added into a culture system of the T cells for culture, so that the efficiently amplified depleted precursor T cells can be obtained, meanwhile, the depletion degree of the CD8 + T cells can be inhibited, and when the prostaglandin PGE1 or PGE2 is stopped, the depleted precursor T cells can be rapidly and directionally differentiated into the CD8 + T cells. Therefore, when the prostaglandin PGE1 or PGE2 is applied to the in-vitro preparation process of adoptive T cell therapy, the total amount of CD8 + T cells can be remarkably increased, the proportion of Tpex is increased, the expression of a depletion marker is reduced, and a technical support is provided for clinical popularization of solid tumor infiltration lymphocyte therapy.

This invention provides methods and compositions for cancer immunotherapy. In various embodiments, the composition comprises functionally enhanced derived effector cells obtained by directed differentiation of genome-engineered iPSCs. In various embodiments, the derived cells provided herein have stable and functional genome editing that delivers improved or enhanced therapeutic effects. Therapeutic compositions and their uses are also provided, comprising these functionally enhanced derived effector cells alone or in combination therapies.

The application discloses a preparation method of a multi-phase "core-shell" biomimetic tissue engineering support, and constructs a multi-phase "core-shell" in-situ biomimetic support to promote the reconstruction of a rotator cuff tendon-bone interface; the support is made of PCL / PEO biological materials and mixed with a plurality of bioactive factor microspheres, the support fiber is a "core-shell" structure, the "shell" contains SDF-1 chitosan slow-release microspheres, the "inner core" is a bone tissue layer, a fibrocartilage layer and a tendon layer, the layers are continuously printed, the spatial distribution of various factors is fine and controllable, the tendon-bone interface is reconstructed in a biomimetic manner, the sequential controllable release of SDF-1 in the outer layer and various differentiation induction factors in the inner core is realized by using the "core-shell" structure fiber, the endogenous MSCs recruitment and the ordered directional differentiation of each layer are promoted, and the application develops an electrohydrodynamic coaxial 3D printing technology on the basis of the electrohydrodynamic printing technology, and constructs a multi-phase "core-shell" in-situ biomimetic support to promote the reconstruction of the rotator cuff tendon-bone interface.

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

The present application relates to the technical field of regenerative medicine application, and particularly relates to a serum-free induction medium with clear components for promoting differentiation of adipose-derived mesenchymal cells into dermal papilla cells, and an induction method and application. A method for promoting differentiation of adipose-derived mesenchymal cells into dermal papilla cells, which is to use a serum-free induction medium to induce culture of adipose-derived mesenchymal cells, so that the adipose-derived mesenchymal cells are differentiated into dermal papilla cells. The formula of the serum-free induction medium comprises hepatocyte growth factor, hydrocortisone, bone morphogenetic protein 2, BIO, and basic fibroblast growth factor. The present application is based on the differentiation potential of adipose-derived mesenchymal cells and the mechanism of maintaining the function of dermal papilla cells, and constructs a serum-free induction system with clear components to realize directional differentiation. The technical scheme can solve the technical problem that there is no method for inducing directional differentiation of adipose-derived mesenchymal cells into dermal papilla cells with effective hair follicle regulation function in the prior art, and has ideal application and popularization potential.

The invention discloses an induction method for directionally differentiating hESC-melanocytes based on a neural crest lineage, and belongs to the technical field of cell engineering and cytobiology. According to the induction method, a complex differentiation process of embryonic stem cells-ectoderm-neural ectoderm-neural crest cells is abandoned, and the neural crest cells are directly obtained from embryonic stem cells, so that the differentiation process is simplified; the differentiation efficiency and controllability of the neural crest cells are remarkably improved, the obtained melanocytes have higher tyrosinase activity and melanin synthesis capacity, and a new way is provided for treating leucoderma by adopting a melanocyte transplantation method.

The invention belongs to the technical field of biological medicine, and particularly discloses injectable hydrogel based on platelet lysate modification, a preparation method of the injectable hydrogel and application of the injectable hydrogel in osteochondral defect repair. The platelet lysate modified injectable hydrogel is photosensitive dual-network hydrogel with a uniform structure, which is constructed by taking platelet lysate (PL), methacrylic anhydride (MA), E7 peptide and polyethylene glycol diacrylate (PEGDA) as raw materials through a photocuring technology, and the hydrogel is cured after in-situ injection at a defect position to promote osteochondral regeneration. The dual-network hydrogel can simultaneously meet the extracellular matrix bionic requirements of cartilage and bone due to the characteristics of rigidity and softness; the combination of the E7 peptide and the PL can effectively utilize a local microenvironment and promote migration and directional differentiation of surrounding stem cells. Therefore, the invention provides a self-adaptive hydrogel scheme which is simple and convenient in preparation process and can promote layered repair of osteochondral defects by using a surrounding matrix environment.

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

This invention relates to the field of biomedical engineering technology and discloses a method for the circular expansion of exosomes from nucleus pulposus cells and its application, comprising the following steps: S1: culturing HUMSCs and NPs separately, collecting the culture supernatant of the NPs and purifying and extracting hNP-exo; S2: diluting the hNP-exo with complete HUMSC culture medium and co-culturing it with in vitro cultured spare HUMSCs to induce the HUMSCs to differentiate into NP-like cells; S3: culturing the NP-like cells, collecting their supernatant and separating and purifying them to obtain NP-like exosomes. This method for the circular expansion of exosomes from nucleus pulposus cells and its application uses human umbilical cord mesenchymal stem cells (HUMSCs) as seed cells to replace traditional bone marrow mesenchymal stem cells (BMSCs). HUMSCs can be obtained from the umbilical cord, which is readily available and can be recycled with the consent of the mother and her family. The acquisition difficulty of HUMSCs is much lower than that of BMSCs, thus solving the defects of BMSCs, which require bone marrow aspiration, involve risks and pain, and have limited collection volume.