572 results about "Organoid" patented technology

The embodiments of the invention described herein relate to systems and methods for culturing and / or maintaining intestinal cells, tissues and / or organoids in vitro. The cells, tissues and / or organoids cultured according to the methods and systems described herein can mimic or reproduce natural intestinal epithelial structures and behavior as well as support co-culture of intestinal microflora.

Culture media and methods for expanding and differentiating populations of stem cells and for obtaining organoids. Expanded cell populations and organoids obtainable by methods of the invention and their use in drug screening, toxicity assays and regenerative medicine.

The invention discloses a method for culturing normal human lung tissue and a lung cancer tissue organoid in an in-vitro manner. The method includes: acquiring fresh human-derived lung tissue cells, and performing collagen digestion on the fresh human-derived lung tissue cells to obtain single cells; culturing human lung tissue and the lung cancer tissue organoid under in-vitro 3D culture conditions; performing H&E staining to determining the structure and form, and using q-PCR to detect related gene expression; using immunofluorescent staining to authenticate cell sources and detect related protein expression. The invention further discloses a method for building a mouse animal model based on the organoid. The method for culturing the normal human lung tissue and the lung cancer tissue organoid and the method for building the mouse animal model have the advantages that the methods are significant to the building of large-scale and good-consistency human-derived in-situ lung cancer animal models, and a good basis and related application prospect are provided for the fundamental researches of lung cancer.

The invention relates to a method for culturing epithelial stem cells, isolated tissue fragments comprising the epithelial stem cells, or adenoma cells, and culturing the cells or fragments in the presence of a Bone Morphogenetic Protein (BMP) inhibitor, a mitogenic growth factor, and a Wnt agonist when culturing epithelial stem cells and isolated tissue fragments. The invention further relates to a cell culture medium comprising a BMP inhibitor, a mitogenic growth factor, and a Wnt agonist, to the use of the culture medium, and to crypt-villus organoids, gastric organoids and pancreatic organoids that are formed in the culture medium.

Culture media and methods for expanding and differentiating populations of stem cells and for obtaining organoids. Expanded cell populations and organoids obtainable by methods of the invention and their use in drug screening, toxicity assays and regenerative medicine.

The invention discloses an in vitro culture method of a colorectal cancer organoid. The in vitro culture method comprises: 1) pretreatment of colorectal cancer biopsy tissue: washing colorectal cancerbiopsy tissue, shearing, and digesting with enzyme to obtain colorectal cancer single cell; and 2) in vitro culture of the colorectal cancer single cell: mixing the colorectal cancer single cell andmatrigel, solidifying, adding a colorectal cancer organoid culture liquid, and culturing to obtain the colorectal cancer organoid. According to the present invention, the method has characteristics ofsimpleness, good stability and high success rate; and the obtained colorectal cancer organoid has a three-dimensional structure, substantially retains the unique morphological characteristics of thepatient tumor tissue, is highly consistent with the pathological features of the biopsy tissue, and provides the material basis for the establishment of the organoid library and the screening of drugs.

The invention relates to a liver organoid, uses thereof and a method for obtaining them.

The invention relates to a method for culturing epithelial stem cells, isolated tissue fragments comprising the epithelial stem cells, or adenoma cells, and culturing the cells or fragments in the presence of a Bone Morphogenetic Protein (BMP) inhibitor, a mitogenic growth factor, and a Wnt agonist when culturing epithelial stem cells and isolated tissue fragments. The invention further relates to a cell culture medium comprising a BMP inhibitor, a mitogenic growth factor, and a Wnt agonist, to the use of the culture medium, and to crypt-villus organoids, gastric organoids and pancreatic organoids that are formed in the culture medium.

The invention relates to the field of culture of tumor organoids, and in particular relates to a special medium used for in-vitro culture of esophagus cancer tumor organoid based on human esophagus cancer tissue and a culture method. The special medium comprises a receptor tyrosine kinase ligand, an Rho-associated coiled-coil protein kinase inhibitor, vitamin and hormone, an antioxidant, and an agonist for inhibiting cell differentiation pathway. According to the method, through the step-by-step culture technology adopting conditional dedifferentiation, tumor cells rapidly amplify within a short time to form the organoid. Through the step-by-step culture technology, primary cells rapidly form the organoid, meanwhile, adequate cells are formed within the effective time, so that the follow-up experiment operation can be conveniently carried out, and therefore, the culture speed and success rate of the organoid are improved.

The invention discloses a lung cancer organ model and an application thereof in tumor research. An establishment method of the lung cancer organ model comprises the steps as follows: (1) lung cancer tumor cells are added to a three-dimensional culture medium for culture, and the lung cancer tumor cells are added to a culture medium to be continuously cultured after a tumor layer is formed, whereinthe culture medium contains growth factors and extracellular matrix suitable for growth lung cancer tumor cells; (2) a system obtained through culture in step (1) forms single cell after protease enzymolysis, resuspension is performed by the culture medium after separation, and then, the lung cancer organ model is obtained. By use of the method, a specimen library containing a large number of lung cancer organ models can be established, heterogeneity and diversity of lung cancer are greatly covered, the defect of insufficient representativeness of traditional lung cancer cell lines is effectively overcome, and the lung cancer organ model becomes an efficient platform for research of occurrence and development mechanisms of lung cancer.

The invention provides a proliferation culture medium and differentiation culture medium for hepatocyte culture and liver organ preparation. The proliferation culture medium and the differentiation medium both take a culture medium for growth of mammalian cells as a basic culture medium, and an agent for supplementing L-glutamine, a pH value modifier for maintaining the pH values of the culture media stable, a primary cell culture antibiotic, a serum substitute, N-acetylcysteine, arbitrary nicotinamide and any one or more of a BMP inhibitor, a Wnt agonist, a growth factor, a Rock signaling pathway inhibitor, a P38 signal path inhibitor, a Notch signal path inhibitor, dexamethasone, BMP7 and a cAMP activator are added into the culture media. By using the culture media for culturing liver cells, functional liver organs can be obtained.

The application relates to a culture medium for nasopharyngeal carcinoma organs, and methods for culturing and identifying the nasopharyngeal carcinoma organs. According to the culture and growth characteristics of nasopharyngeal carcinoma-derived cells, a variety of cytokine components are selected and blended according to a certain ratio; the contents of cytokines and signal pathway regulatory factors in the culture medium are suitable; the nasopharyngeal carcinoma cells can effectively form organoids in a three-dimensional (3D) environment. The methods for culturing and identifying the nasopharyngeal carcinoma organs as well as the unique culture medium for the nasopharyngeal carcinoma organs can be used for successfully and stably culturing the nasopharyngeal carcinoma organs which have pathophysiological features and genotypes being highly consistent with those of the parental tumor and have high similar form of organization.

Methods of tissue engineering, and more particularly methods and compositions for generating various vascularized 3D tissues, such as 3D vascularized embryoid bodies and organoids are described. Certain embodiments relate to a method of generating functional human tissue, the method comprising embedding an embryoid body or organoid in a tissue construct comprising a first vascular network and a second vascular network, each vascular network comprising one or more interconnected vascular channels; exposing the embryoid body or organoid to one or more biological agents, a biological agent gradient, a pressure, and / or an oxygen tension gradient, thereby inducing angiogenesis of capillary vessels to and / or from the embryoid body or organoid; and vascularizing the embryoid body or organoid, the capillary vessels connecting the first vascular network to the second vascular network, thereby creating a single vascular network and a perfusable tissue structure.

The invention discloses a three dimensional tissue and organ culture model, a high throughput automatic stereo image analyzing platform thereof, and a method utilizing the provided platform to screen antitumor drugs. The platform can be applied to the culture system of three dimensional tissue and organ / organ-like substance, three dimensional scanning and sampling system, and high-volume data storage and computer analysis system. The provided platform can simultaneously process a large amount of clinical samples and screen a plurality of antitumor drugs, is capable of greatly reducing the cost and maximally reducing the detection time, thus is widely applied to the drug selection schemes and dosage optimization in clinic, novel drug development, and basic researches on interactions among tissue, organ, biological macromolecules, and other micromolecules.

The invention discloses a specific culture medium for a lung tumor organ and a stentless 3D culturing method. The specific culture medium is prepared from the following components: FBS, double antibody, N-2, Noggin, B-27, EGF, FGF-10, Y-27632, A 83-01, SB202190, N-acetylcysteine, HEPES, Glutamax, IGF-1, hydrocortisone and Advanced DMEM / F12. The culturing method comprises the following steps: adding a tumor cell into a low serum culture medium, re-suspending the tumor cell, inoculating the tumor cell into a culture vessel, adding the specific culture medium into the culture vessel, changing thespecific culture medium once a day, and performing culturing until an organoid is formed. According to the culture medium and culturing method, a tumor organoid can quickly generate, can be stably cultured for a long time, is regular in spheroid form and has uniform and controllable size, and the heterogeneity of a tumor tissue of a patient can be well maintained in vitro.

The invention discloses a method for separation and in-vitro culture of pancreatic cancer tissue organs of human. The method comprises the following steps that (1) after the pancreatic cancer tissue of human is cut, enzymic digestion is repeatedly carried out for 10-20 minutes at 37 DEG C in a rotation speed of 30-40 rpm three times, and a pancreatic cancer single cell is obtained; (2) the pancreatic cancer single cell and matrigel are uniformly mixed, then a culture medium containing a growth factor PEG2 and gastrin I are added for culture, and the pancreatic cancer tissue organs of human areobtained. The method for separation and in-vitro culture of the pancreatic cancer tissue organs of human has the advantages that the tumor tissue is digested by adopting a short-time repeated digestion method, which can effectively improve the cell yield and cell viability; the culture medium containing the growth factor PEG2 and the gastrin I is adopted for culture, which can effectively promotethe growth of the pancreatic cancer organs and increase the survival rate of the pancreatic cancer organs.

An in vitro liver organoid is provided along with methods of making and using the organoid. The liver organoid can be manipulated to have either immature or mature characteristics. Some manipulations generate a liver organoid that supports expansion and / or differentiation of hematopoietic stem cells.

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.

The invention provides a preparation method of organoid spheres. The preparation method comprises the steps that matrigel containing cells and fluorocarbon oil are introduced into a three-way device to obtain cell spheres, and after culture is performed, the organoid spheres are formed; according to the preparation method of the organoid spheres, the micro-fluid control method is adopted for generating mono-disperse biomaterial organs, high-throughput production is achieved, the sizes, shapes and structures of organoids are controllable and uniform, and compared with a 2D tumor sensitivity predicting model, the organoids are combined with tumor micro-environment factors, and the prediction result is more accurate; compared with a PDX mouse tumor sensitivity predicting model, the modeling period is shorter, and the cost is lower; compared with common gene sequencing, the clinic predicting rate is higher; the preparation method of organoid spheres has the important significance and valuefor clinic application, and the important basis is supplied to detection of related results.

Described are cell culture solutions and systems for epithelial stem cell and organoid cultures, formation of epithelial constructs and uses of the same in transplantation. A single layer of epithelial cells that actively self-renews and is organized into crypts and villi clothes the intestine. It has been recently shown that the renewal of intestinal epithelium is driven by Lgr5+ intestinal stem cells (ISC) that reside at the base of these crypts (Barker et al., 2007). Lgr5+ stem cells can be isolated and cultured in vitro to form organoids containing crypt-vcllus structures that recapitulates the native intestinal epithelium (Sato et al., 2009).

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 al 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.

The invention provides an in vitro culture method of cancer type organs. According to the embodiment of the invention, the method comprises the steps of performing shearing and digestion treatment oncancer tissue samples, wherein the size of cancer tissue after the shearing treatment is 1-2mm<3>, and the digestion treatment is performed under the action of combined digestive enzymes; and performing re-suspension treatment on cancer cells obtained after digestion treatment with precooling substrate glue, wherein the concentration of the cancer cells in the substrate glue is 500-1500 / 10 [mu]L;and performing point and pallet treatment on cancer cell suspension, and performing culture treatment on the cancer cells after point and pallet treatment is performed, so as to obtain cancer organs.According to the method of the embodiment of the invention, the enzymolysis efficiency of the cancer cells is improved, the success rate of subculturing of organoids is increased, and long-term culture of the organoids can be realized, and a biology sample database can be established.

The invention discloses a 3D culture, passage, cryopreservation, recovery and identification method for in-vitro organoids based on small intestines of different segments of mice. The method comprisesthe following steps: (1) separating recesses of duodenum, jejunum and ileum segments of mice; (2) performing 3D culture on the recesses of the duodenum, jejunum and ileum segments of mice, and forming organoids; (3) performing passage on organoids of small intestines of mice; (4) performing cryopreservation on the organoids of small intestines of mice; (5) performing recovery on the organoids ofsmall intestines of mice; (6) preparing frozen sections of the organoids of small intestines of mice; and (7) performing immunofluorescence labeling and identification on the frozen sections of the organoids of small intestines of mice. Compared with the prior art, the method disclosed by the invention optimizes a recess extraction manner, an in-vitro culture system and a culture medium based on the recesses of the small intestines containing stem cells, and comprises the steps of complete culture, passage, cryopreservation, recovery and identification. The small intestine organoids obtained by the method are capable of performing repeated passage, and the passage organoids have character stability.

The invention relates to the field of accurate medical treatment and cell biology, in particular to a culture method of an in-vitro substance used for scientific research experiments. The method comprises the following steps: 1, treatment of breast cancer tissues from patients: (1) treating fresh breast cancer sample tissues; (2) performing collagenase digestion after shearing; and (3) acquiring breast cancer primary cells; and 2, in-vitro culture of the breast cancer primary cells: (1) mixing the breast cancer primary cells with an organoid culture medium; (2) after solidification, adding thesolidified mixture into a breast cancer organoid culture solution for culture; and (3) acquiring a breast cancer organoid.

The invention discloses a contour perception multi-organ segmentation network construction method based on class-by-class convolution operation. The contour perception multi-organ segmentation networkconstruction method comprises the following steps: 1, performing region coarse segmentation and edge detection on multiple organs of the abdomen; 2, introducing a semantic-guided class-by-class multi-scale attention mechanism; step 3, performing class-by-class fusion of multi-branch information; step 4, performing introduction of multi-task loss. According to the invention, the advantages of a convolutional neural network and a gated recurrent neural unit are utilized, and for the characteristics and difficulties of a multi-organ segmentation task, via the contour information assisted multi-scale feature extraction, a class-by-class multi-scale semantic attention mechanism, a class-by-class cavity convolution fusion mechanism and a plurality of loss functions can be introduced to relievethe inter-class imbalance problem of organs; multi-organ segmentation is performed on a three-dimensional CT image more efficiently and accurately, and the advantages of the invention are verified ona data set containing 14 types of organ labels; the invention can be widely applied to computer-aided diagnosis and treatment application, such as endoscopic surgery, interventional therapy and radiotherapy plan making.

Described herein are organoids comprising admixtures of selected bioactive primary renal cells and a bioactive cell population, e.g., an endothelial cell populations, e.g. HUVEC cells, and methods of treating a subject in need thereof with such organoids. Further, the isolated renal cells, which may include tubular and erythropoietin {EPO}-producing kidney cell populations, and / or the endothelial cell populations may be of autologous, syngeneic, allogeneic or xenogeneic origin, or any combination thereof. Further provided are methods of treating a subject in need with the organoids.

The invention discloses a culture medium and an organoid culture method for 3D culture of kidney tissue organoids. According to culture and growth characteristics of kidney-tissue-derived cells, the culture medium disclosed by the invention is prepared by blending a plurality of selected cytokine components in certain proportion; so that, signaling pathway regulatory factor content of the cytokines in the blended culture medium is appropriate. Thus, effective formation of organoids by kidney cells and renal cancer cells in 3D environment is allowed.