44 results about "Epithelial Stem Cell" patented technology

The invention discloses a mouse lung organ and fibroblast co-culture method, which comprises the following steps of: performing in-vitro mouse lung organ culture on mouse lung tissue primary epithelial stem cell spheres (pneumospheroids) in a Transwell upper chamber, and after the lung organs in the Transwell upper chamber are differentiated, replacing with a lower chamber paved with mouse lung fibroblasts, therefore, a gas-liquid interface co-culture system of the mouse lung organ (upper chamber) and the fibroblast (lower chamber) is formed, and the hierarchical structure, morphological characteristics and regeneration function of the upper chamber organ and the phenotype, function and related signal path activation level of the lower chamber fibroblast can be dynamically researched.

The invention discloses an amniotic epithelial stem cell screening method and device based on microscopic image feature extraction, relates to the technical field of microscopic image analysis and cell screening, and is used for solving the problem that group topology and internal activity in amniotic epithelial stem cell lossless screening are difficult to collaboratively quantify. The method comprises the following steps: firstly, performing enhancement processing on a phase difference microscopic image, and extracting a kernel centroid and a global gradient energy diagram; then, constructing a Voronoi jurisdiction, and coupling a cytoplasm texture entropy and a gray scale attenuation gradient to generate a cytoplasm compactness factor; meanwhile, calculating a cell gap topology consistency index based on mapping of the Voronoi boundary and the gradient map; and finally, weighting and correcting the compactness factor by using a topological index to obtain a dryness maintenance confidence coefficient, and outputting a target coordinate. Through dual verification of geometric topology and physical gradient, combined constraint screening of internal and external characteristics of cells is realized.

Methods and systems are described for a high-level approach to organ generation by self-assembly using bioprinting, overcoming longstanding challenges in replicating the intricate architecture and function of complex organs. In some embodiments, these methods are used to produce lung tissues and microtissues, demonstrating how mini-lung constructs can be produced on a small scale through specialized processes and media. In such embodiments, methods and systems for generating bioprinted lung tissues and microtissues are provided, as are tissues made by such methods. The tissues find use in implantation, drug discovery, personalized medicine, and other applications. Methods for the large-scale manufacturing of lung epithelial stem cells are also provided.

The application discloses an amnion epithelial stem cell screening method and device based on microscopic image feature extraction, relates to the technical field of microscopic image analysis and cell screening, and is used for solving the problem that the population topology and internal activity are difficult to be quantified in the lossless screening of amnion epithelial stem cells. Firstly, phase contrast microscopic images are subjected to enhancement processing, and nucleolus centers and global gradient energy maps are extracted; subsequently, Voronoi districts are constructed, and cytoplasm texture entropy and gray attenuation gradient are coupled to generate a cytoplasm compactness factor; meanwhile, a cell gap topology consistency index is calculated based on the mapping of the Voronoi boundary and the gradient map; finally, the compactness factor is weighted and corrected by using the topology index to obtain stemness maintenance confidence, and target coordinates are output. Through the dual verification of geometric topology and physical gradient, the application realizes the joint constraint screening of the features inside and outside the cells.

Methods and compositions for treating a corneal wound and / or increasing a population of limbal epithelial stem cells (LESCs) and / or early transit amplifying cells (eTAs) in the limbal epithelium of a subject in need thereof. The methods can include administering a therapeutic agent to the subject that: i) results in an increase in concentration of IFITM1 (Interferon Induced transmembrane Protein 1) in the limbal epithelium of the subject; and / or ii) results in a decrease in concentration of OVOL1 (Ovo Like Zinc Finger 1) in the limbal epithelium of the subject. Also provided are adeno-associated viruses (AAVs). The AAVs comprise a heterologous nucleic acid sequence encoding IFITM1 or a variant thereof, operably linked to a regulatory sequence.

The application provides an amniotic membrane stem cell and a separation method and a hydrogel 3D culture method thereof, relates to the technical field of cell culture, and comprises the following steps: placing amniotic membrane fragments in a pre-digestion buffer in sequence and incubating for 10-20 min, placing the amniotic membrane fragments in a mixed enzyme solution and performing digestion treatment, filtering, and obtaining a filtrate and residual amniotic membrane fragments; centrifuging the filtrate to obtain a cell precipitate, and performing differential adhesion culture to obtain amniotic membrane epithelial stem cells; adding a culture medium to resuspend the residual amniotic membrane fragments so that the residual amniotic membrane fragments are in contact with a culture container and are in a non-suspended state, performing adhesion culture, and obtaining amniotic membrane mesenchymal stem cells that have climbed out. Compared with the number of cells and the cell survival rate of cells that are subjected to digestion treatment of other mixed enzymes, the number of cells obtained by digestion treatment of the mixed enzyme is significantly increased, and the cell survival rate is higher; compared with a step-by-step digestion strategy, the mixed enzyme digestion one-step method is more simple, and efficient and synchronous separation is achieved. The technical problem of a significant decrease in cell activity caused by the step-by-step digestion separation strategy in the prior art is solved.

Provided herein are constructs of micro-aggregate multicellular, minimally polarized grafts containing Leucine-rich repeat-containing G-protein coupled Receptor (LGR) expressing cells for wound therapy applications, tissue engineering, cell therapy applications, regenerative medicine applications, medical / therapeutic applications, tissue healing applications, immune therapy applications, and tissue transplant therapy applications which preferably are associated with a delivery vector / substrate / support / scaffold for direct application.

An artificial hair follicle germ 10 has a three-dimensional structure in which a first block 11 including hair follicle epithelial stem cells, a second block 12 including hair papilla cells, and a third block 13 including dermal-derived mesenchymal cells (DDMCs) are stacked in this order. A method for artificially preparing a hair follicle germ 10 includes three-dimensionally culturing a first block 11 including hair follicle epithelial stem cells, a second block 12 including hair papilla cells, and a third block 13 including dermal-derived mesenchymal cells (DDMCs), these blocks being stacked in this order.

To provide an external composition that stably contains minoxidil in high concentration, and exhibits better hair growth effect than that of existing treatment agents.SOLUTION: An external composition contains 2 w / v% or more of minoxidil, the minoxidil being fine particles with an average particle size of 50 nm-200 nm. The external composition preferably further contains cellulosic polymers.SELECTED DRAWING: Figure 3

The invention provides a method for constructing a tissue engineered cornea based on human amniotic epithelial stem cells and application, and belongs to the technical field of biology. The method comprises the following steps: 1) culturing human amniotic epithelial stem cells in a corneal epithelial cell induced differentiation culture medium to obtain human corneal epithelial cell-like cells; culturing the human amniotic epithelial stem cells in a corneal stroma cell induced differentiation culture medium to obtain corneal stroma cells; and 2) respectively inoculating the human corneal epithelial cell-like cells and corneal stroma cells obtained in the step 1) on the surface and inside of an acellular corneal stroma scaffold, and co-culturing for 5 days to construct the tissue engineered cornea. According to the method, the two corneal cells derived from the human amniotic epithelial stem cells are simultaneously inoculated to the human decellularized corneal stroma for the first time, the tissue-engineered cornea is successfully constructed, in addition, the corneal cells obtained through differentiation can be inoculated to various types of biological materials such as hydrogel, and the tissue-engineered cornea is obtained. Wider application possibility is provided for developing various novel artificial cornea products.

The present invention relates to functionally improved epithelial stem cells. The epithelial stem cells with improved functionality by a method of the present invention are prepared without genetic modification, thus being less likely to cause side effects due to DNA delivery / modification, are derived from the amniotic membrane in which no tissue-compatible antigen (factor) is expressed, thus having no rejection, and exhibit improved cell migration and would healing effects due to the improved functionality. Therefore, the epithelial stem cells can be used for the regeneration of various tissues, including the cornea, amniotic membrane, skin, and the like, and particularly can be used for the prevention or treatment of premature rupture of the amniotic membrane.

To develop a novel method for treatment of diseases leading to serious corneal and conjunctival epithelial damage resulting from functional impairment of corneal and conjunctival epithelial stem cells, including severe ocular surface disease attributable to chronic graft-versus-host disease.SOLUTION: The present invention provides a prophylactic or treatment agent for severe ocular surface disease for topical administration, comprising apocynin as an active ingredient.SELECTED DRAWING: None

The purpose of the present invention is to develop a novel treatment method for diseases such as severe ocular surface diseases caused by chronic graft-versus-host disease, which cause severe corneal and conjunctival epithelial damage due to dysfunction of corneal and conjunctival epithelial stem cells. SOLUTION: The present invention provides a topically administered agent for preventing or treating severe ocular surface diseases, which contains apocynin as an active ingredient.

The application discloses application of human amniotic epithelial stem cells in preparation of a medicine for treating depressive diseases. The human amniotic epithelial stem cells are obtained by mechanically separating the inner surface amnion of discarded placental tissue, and then subjected to trypsin digestion, centrifugation, resuspension and harvesting in sequence after washing; and a human amniotic epithelial stem cell scheme for treating depression; the cells are widely sourced, free of ethical controversy, low in immunogenicity and non-tumorigenic, and provide a new safe and effective approach for treating depression; the cell preparation can be administered through intravenous injection, intrathecal injection or nasal delivery, and the mechanism of action is to inhibit the overactivation of glial cells, reduce the depressive-related neuroinflammatory response, and thus improve the neuronal microenvironment. The application can effectively promote hippocampal neurogenesis (increase in DCX positive cells), regulate 5-hydroxytryptaminergic neurons and glutamatergic neuron activity, restore the steady state of neural networks, and significantly improve the depressive behavior of model mice.

To provide a defined culture medium for isolating stratified epithelial stem cells and stably maintaining the epigenetics of the stratified epithelial stem cells during multiple passages, and to provide clonal stratified epithelial stem cells isolated using the defined culture medium.SOLUTION: And a defined culture medium comprising each of ROCK (Rho kinase) inhibitors, mitogenic growth factors, insulin or IGF, TrkA inhibitors, and Oct4 activators, and comprising at least one of VEGF inhibitors, tyrosine kinase inhibitors, and / or FGF10 or FGF10 agonists, and optionally further comprising TGF β signaling pathway inhibitors and / or bone morphogenetic proteins (BMP) antagonists, wherein the defined culture medium supports epigenetically stable growth and proliferation of stem cells of stratified epithelial tissue origin in culture.SELECTED DRAWING: Figure 1

The invention discloses a mouse airway-like and fibroblast co-culture method, which comprises the following steps: carrying out in-vitro mouse airway-like culture on lung epithelial stem cells dissociated from mouse lung tissue primary epithelial stem cell spheres (pneumospheroids) in a Transwell upper chamber, after airway-like differentiation of the Transwell upper chamber is completed, replacing the Transwell upper chamber with a lower chamber paved with mouse lung fibroblasts, and carrying out in-vitro mouse airway-like culture. During co-culture, only a culture medium is added into the lower chamber, and the culture medium adopts the same culture medium (gas-liquid interface differential culture medium) as the culture medium in the airway-like culture process, so that a gas-liquid interface co-culture system of the mouse airway-like (upper chamber) and lung fibroblasts (lower chamber) is formed, and the hierarchical structure, morphological characteristics and regeneration function of the airway-like of the upper chamber can be dynamically researched; and lower ventricle fibroblast phenotypes, functions, and related signaling pathway activation levels.

The application provides a method for constructing a tissue-engineered cornea based on human amniotic epithelial stem cells and application thereof, and belongs to the technical field of biotechnology. The method comprises the following steps: 1) culturing human amniotic epithelial stem cells in a corneal epithelial cell induction and differentiation culture medium to obtain human corneal epithelial cell-like cells; culturing the human amniotic epithelial stem cells in a corneal stromal cell induction and differentiation culture medium to obtain corneal stromal cells; 2) inoculating the human corneal epithelial cell-like cells and the corneal stromal cells obtained in step 1) on the surface and inside of a decellularized corneal stromal scaffold respectively, and co-culturing for 5 days to construct a tissue-engineered cornea. The application successfully constructs a tissue-engineered cornea by inoculating two kinds of corneal cells derived from human amniotic epithelial stem cells on a human decellularized corneal stromal at the same time for the first time. In addition, the obtained corneal cells can be inoculated on various types of biomaterials such as hydrogels, thereby providing a wider application possibility for developing various types of artificial cornea products.

The purpose of the present invention is to develop a novel method for treating diseases leading to severe corneal and conjunctival epithelial disorder from dysfunction of corneal / conjunctival epithelial stem cells, such as severe ocular surface diseases caused by chronic graft-versus-host disease.　The present invention provides a prophylactic or therapeutic agent for severe ocular surface disease for topical administration, the agent containing apocynin as an active ingredient.

The application discloses application of TACSTD2 protein in preparation of products for detecting and identifying human amniotic epithelial cells, and belongs to the technical field of biological medicines. The application proposes a protein marker for detecting and identifying human amniotic epithelial stem cells, which comprises TACSTD2 protein, or TACSTD2 protein combined with one or more selected from CD29, CD166, CD73, CD90, CD105, CD324, CD326, SSEA4, Nestin and Vimentin. The application further proposes application of a reagent for detecting TACSTD2 protein expression in preparation of products for detecting and identifying human amniotic epithelial cells, which is specifically highly expressed in human amniotic epithelial stem cells and can be used alone or in combination with other cells for detecting and identifying human amniotic epithelial cells.

This invention discloses the application of genes derived from the rumen abdominal sac tissue of ruminants in promoting the differentiation of rumen epithelial stem cells, belonging to the field of genetic engineering technology. This invention is the first to discover genes with unknown functions. ABCC1、NPC1L1 , MRPL24 or GPR15LG The proteins encoded by these substances promote the differentiation of rumen epithelial stem cells, and this can be achieved by regulating gene expression or utilizing their encoded proteins to regulate the proliferation and differentiation of rumen epithelial stem cells. The research results of this invention overcome the limitations of traditional breeding methods that rely on phenotypic selection, by screening… ABCC1、NPC1L1 , MRPL24 or GPR15LG This study provides a reference for individuals with high gene expression and for the targeted breeding of breeds with more efficient rumen function using biological methods, and provides a core target for the breeding of new ruminant breeds that are high-yielding, efficient, and stress-resistant.

This invention relates to the application of human amniotic epithelial stem cells (hAESCs) in retinal regeneration and repair, belonging to the field of biomedical technology. It aims to address the technical problems of off-target effects, low efficiency, and safety risks associated with existing methods that directly reprogram Müller glial cells (MGs) via viruses. This invention transplants hAESCs into the degenerated subretinal space. hAESCs, through secreting active substances such as matrix metalloproteinases (MMPs), degrade excessively deposited extracellular matrix, improve the pathological microenvironment inhibiting regeneration, and thus promote the reprogramming of endogenous MGs into functional retinal neurons, thereby achieving retinal structural repair and visual function restoration. This invention utilizes highly safe hAESCs and, by remodeling the microenvironment, significantly improves the efficiency and specificity of MG differentiation into functional neurons, providing a new strategy for the treatment of retinal degenerative diseases.

To provide a method for isolating normal stem cells in a culture derived from columnar epithelial tissues and cancer stem cells derived from epithelial cancers, and to provide a culture media system useful for epigenetically stable propagation.SOLUTION: Provided is a method for isolating a stem cell from epithelial tissue, preferably columnar epithelial tissue, comprising: (i) culturing dissociated epithelial cells derived from a columnar epithelial tissue sample to form stem cell colonies; (ii) isolating single stem cells from the cell colonies; and (iii) culturing isolated single stem cells from step (ii) individually to form cultures purified stem cell clones, each of the stem cell clones representing a clonal expansion of an epithelial stem cell present in the columnar epithelial tissue sample, thereby isolating epithelial stem cells.SELECTED DRAWING: None

Epithelial stem cells (15), such as hair follicle epithelial stem cells, are cultured on a scaffold (13) comprising a molecule containing an extracellular cadherin (cadherin EC) domain. The epithelial stem cells 15 are then separated from the scaffold 13. The epithelial stem cells 15 are prepared through the abovementioned process. Thereafter, a hair follicle primordium10 may be produced by three-dimensionally culturing the epithelial stem cells 15 and mesenchymal cells 16 in a state in which a layer of the epithelial stem cells 15 and a layer of the mesenchymal cells 16 overlap. The mesenchymal cell 16 layer may include a layer of hair papilla cells and a layer of cells constituted of dermal pluripotent cells (DDMCs) or dermal sheath cells (DS cells). The epithelial stem cell 15 layer may overlap the hair papilla cell-layer side of the mesenchymal cell 16 layer.

The invention relates to cell biology and regenerative medicine, specifically to a culture method and application for porcine limbal epithelial stem cells. The method involves pretreatment of cells, coating of culture dishes, centrifugation, culturing, passaging, and identification by immunofluorescence staining. The cultured cells are applicable in repairing corneal diseases, including limbal stem cell deficiency. During repair, the cells can be combined with biological materials to form a stem cell engineering carrier for implantation at the injury site. The separation and culture method is simple, yields a large number of cells, and is suitable for use with porcine eyeballs. Limbal epithelial stem cells cultured in vitro form distinct cell clusters, referred to as stem cell islands. The obtained cells exhibit phenotypes including BMI1+, ΔNp63α+, CK14+, and ABCG2+. Based on experimental and clinical research, this method provides a promising approach for cell transplantation in ocular surface reconstruction.

The application discloses a radial expansion type flexible clamping and stacking device for amniotic epithelial stem cell culture dishes, and belongs to the technical field of cell culture engineering. The radial expansion type flexible clamping mechanism can greatly reduce the interference to amniotic epithelial stem cells during clamping of the culture dishes, the design can significantly reduce the displacement amplitude of the culture dishes compared with the large-area adhesion clamping of the traditional clamping mechanism, and effectively prevents uneven distribution of amniotic epithelial cells or falling of the amniotic epithelial cells from the bottom of the culture dishes caused by impact and friction.

Provided herein include methods, compositions, isolated cells, cell models, and kits suitable for use in profiling various cell types in amnion including amnion epithelial cells, amnion mesenchymal cells, fibroblasts, macrophages, as well as amnion stem cells including amnion epithelial stem cells and amnion mesenchymal stem cells. The present disclosure also discloses novel markers for various cell types in human amnion, including marker genes, transcription factors and signaling molecules associated with various stages of amniotic development. The methods, cell models, and gene signatures disclosed herein can be used for a variety of research, diagnostic, or therapeutic applications.

The invention relates to differentiation methods for progenitor cells, e.g. mammalian epithelial stem cells, differentiation media for use in said methods, organoids and cells obtainable by said methods and uses, including therapeutic uses, thereof.

The present application relates to the field of cell detection and screening, and particularly relates to a method and system for detection and screening of amniotic epithelial stem cells. The method comprises: generating a structured sample task sheet based on multi-source information; performing tissue digestion processing based on the task sheet to obtain a candidate cell group; then performing phenotype detection, molecular detection and quality detection to generate corresponding data packets and label data packets; based on the quality label data packets, performing gating, batch normalization, abnormality rejection, fusion scoring and consistency determination processing through a screening engine to generate a screening result packet; and finally generating a disposal work order and a traceability report based on the screening result packet. The present application realizes standardized and auditable automatic processing of the whole process of amniotic epithelial stem cells from receiving, detection to sorting and disposal, greatly improving the detection efficiency, result consistency and traceability.