167 results about "Organ culture" patented technology

A device for performing a biological modification of a fluid, the device includes (a) a chamber having an inlet for intake of the fluid and an outlet for outflow of the fluid; and (b) a collection of micro-organ cultures of at least one organ for performing the biological modification of the fluid, each individual micro-organ culture of the collection including cells and having dimensions, such that cells positioned deepest within the individual micro-organ culture are at least about 100 micrometers and not more than about 225 micrometers away from a nearest surface of the individual micro-organ culture, thereby in vivo organ architecture (organ structure) of organ units (e.g., acinus of liver) is maintained within each individual micro-organ culture, the collection of micro-organ cultures being located within the chamber and the collection of micro-organ cultures being in contact with at least a portion of the fluid flowing through the chamber.

A device for performing a biological modification of a fluid, the device includes (a) a chamber having an inlet for intake of the fluid and an outlet for outflow of the fluid; and (b) a collection of micro-organ cultures of at least one organ for performing the biological modification of the fluid, each individual micro-organ culture of the collection including cells and having dimensions, such that cells positioned deepest within the individual micro-organ culture are at least about 100 micrometers and not more than about 225 micrometers away from a nearest surface of the individual micro-organ culture, thereby in vivo organ architecture (organ structure) of organ units (e.g., acinus of liver) is maintained within each individual micro-organ culture, the collection of micro-organ cultures being located within the chamber and the collection of micro-organ cultures being in contact with at least a portion of the fluid flowing through the chamber.

The present invention relates to in vitro cultured skin substitutes, and in particular to in vitro cultured skin substitutes that have improved barrier function. In some embodiments, improved barrier function is a result of improved culture conditions, while in other embodiments, improved barrier function results from genetic modification of keratinocytes. Improved culture conditions to improve barrier function include organotypic culture in the presence of linoleic acid and / or linoleic acid at about 75% humidity. Suitable genetic modifications for improving barrier function includes transfection with a DNA construct capable of expressing GKLF.

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.

Disclosed are an ophthalmic lens, a cell or organ culture substrate, a container for a biological material and a transparent gel which have excellent wetting and sticking properties and do not require any surface treatment. An ophthalmic lens, cell or organ culture substrate, container for a biological material or transparent gel produced by polymerizing a cyclic siloxane compound represented by the formula (A): (A) wherein Ra are Rb independently represent a hydrogen or a monovalent hydrocarbon group which may be substituted by a fluorine; Rc represents an alkyl group having 1 to 6 carbon atoms or a phenyl group; X represents an organic group having an aliphatic unsaturated bond therein; and n is an integer of 1 to 10.

The invention discloses a three-dimensional cell culture system, comprising container, and cell culturing medium and three- dimentioal cell culture unit inside container; said three- dimentional cell culture unit comprises implanting cell and cavity flavoable for cell adsoption, growth, differentiation and maturity. Said three- dimentional culture unit is transparent, which is flavouable for obeservation of adsorption, enlargement, transition, proliferation, differentiation, maturity, aging, death and tissue or organ development. The invention can be used for long term cell culture and large amount of cell production as seed cell for tissue engineering, and for short term regenerative tissue and micro organ formation for replanting. The invention provides special condition for tumor external research, and convenient for tumor diagnosis, observation of tumor immersion and transfer, and anti- tumor medicine sift. It can aslo be used for cell isolated culture and for research of gene programm change and influence to cell from rxternal environment.

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 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 provides a hair follicle stem cell separation culture method. The method utilizes an organ culture method, adopts the William's E complete medium to culture the hair follicle, separate and purify to get hair follicle stem cells. The method only uses very small skin materials, the rat whisker follicle is separated with a stereoscope through a micro separation method, then the whole whisker follicle is cultured, the hair follicle stem cells grow out from the projection part and form clone, so the cell clone forming efficiency is high, the purification is easy, long-term passage can be realized, the characteristics of the hair follicle stem cells are kept unchanged, and only one whisker follicle can obtain enough hair follicle stem cells for long-term use.

The invention discloses a method for culturing bladder cancer organs in vitro. The method comprises the following steps: manufacturing a gas-liquid interaction culture system, namely manufacturing a uniformly paved rat tail collagen supporting layer on the surface of a porous culture membrane in a Transwell upper chamber; re-suspending fresh in-vitro bladder cancer tissues by using a rat tail collagen solution, uniformly mixing, adding the obtained mixture onto the rat tail collagen supporting layer, and then putting the rat tail collagen supporting layer into an incubator at 37 DEG C to solidify so as to obtain a rat tail collagen layer containing the bladder cancer tissues; adding an organoid culture medium into a Transwell lower chamber, and enabling the liquid level of the culture medium to be lower than the rat tail collagen layer containing bladder cancer tissues; and carrying out passage and cryopreservation. The culture success rate of the bladder cancer organoid is remarkablyincreased, the bladder cancer organoid with reserved immune cells can be obtained through culture, operation is easy, the utilization rate of tumor tissue is high, and great significance and value areachieved for bladder cancer drug screening research.

An agent for dehydrating corneas, in particular eye corneas for organ culture, contains the culture medium and as dehydrating substance hydroxyethylstarch with a mean molecular weight Mw from 070,000 to 200,000, an MS substitution degree from 0.15 to 0.5, a DS substitution degree from 0.15 to 0.5 and a C2-C6 substitution ratio at the anhydroglucose units < / =8. The dehydrating substance is preferably hydroxyethylstarch with a mean molecular weight Mw 130,000+ / -20,000, an MS substitution degree from 0.38 to 0.45, a DS substitution degree from 0.32 to 0.40 and a C2-C6 substitution ratio at the anhydroglucose units from 8 to 20. The hydroxyethylstarch is used in a concentration from 1 to 20 (weight / vol.) %, preferably from 2 to 15 (weight / vol.) %, and in particular of 7.5% (weight / vol.). Also disclosed is the use of this hydroxyethylstarch as a dehydrating substance for the organ culture of corneas, in particular eye corneas, and the use of this hydroxyethylstarch for preparing a dehydrating agent for corneas, in particular eye corneas for organ culture.

The invention relates to the fields of cytobiology and cell culture, and particularly relates to a breast cancer organoid culture kit. A method for realizing the kit comprises the following steps: (1)preparing, sub-packaging and using a breast cancer organoid culture medium A; (2) preparing, sub-packaging and using a breast cancer organoid culture medium B; and (3) preparing, sub-packaging and using a sample preserving fluid. The breast cancer organoid culture medium A is a DMEM / F12 culture medium containing 100 U / ml of penicillin, 0.1 mg / ml of streptomycin, 1% of HEPES and 1% of Gluta Max 100 X; the organoid culture medium B is a DMEM / F12 culture medium containing EGF, an FGF growth factor, an ALK inhibitor A83-01, Noggin, an HEPES buffer solution, Y27632 and one or more of a Wnt signalpath related secretory protein family R-Spondin 1-4; and the sample preserving fluid is a DMEM / F12 culture medium containing 100 U / ml of penicillin, 0.1 mg / ml of streptomycin and 1% of L-glutamine. The three components form the breast cancer organoid culture kit.

The present invention provides a culture medium and a culture method for colorectal cancer organoids. The culture medium comprises a basic culture medium Advanced DMEM / F12, specific additive factors and sterile water; wherein a mass ratio of the basic culture medium Advanced DMEM / F12 to the sterile water is 99:1; and the specific additive factors comprise B27 without vitamin A, N-acetylcysteine, EGF, Noggin, R-spondin 1, Wnt3a, CHIR99021, thiazovivin, Gastrin I, a penicillin-streptomycin mixture and Primocin. The culture medium is used to culture the colorectal cancer organoids, can maintain morphological structures and genetic characteristics of primary tissues, effectively reduces risks of microbial contamination in colorectal cancer culture, and improves success rate and survival rate of colorectal cancer organoid culture.

The invention relates to a culture method of breast cancer organoids and a co-culture method of tumor-related fibroblasts. According to the invention, a more efficient and convenient improved system for key technical links such as PDO culture, passage, maintenance and the like of tumor tissue organoids of breast cancer patients is established, operation steps and methods for establishing and passage of a PDO cell line are simplified in multiple aspects, components of a culture medium, digestive juice, a washing solution and the like are optimized, and finally, a breast cancer PDO culture system is successfully established. Compared with the prior art, the culture method has the advantages that the success rate and the passage frequency are basically consistent, but the operation is simpler, the required time is shorter, a good foundation is laid for a PDO model to have better clinical and scientific research applications in the future, and the corresponding cost is saved. The inventionalso establishes a stable co-culture system of breast cancer PDO and tumor-associated fibroblasts (CAFs), simultaneous culture, amplification and passage of PDO and CAFs can be realized, and the PDOand the CAFs do not produce obvious influence on each other.

The invention relates to asparagus parent breeding method, mainly solving the problems that rooting rate is low, root quality is poor and the survive rate of the regeneration plant is low in asparagus organ tissue culture process. The method includes the process that high quality asparagus stem section is selected, culture is carried out in culture medium solution, and asparagus seedling meeting transplant standard is bred. The culture medium solution is confected by the way that hormones corresponding to various culture stages are added on the basis of the basic culture medium solution, the culture includes induction culture, propagation culture, rooting pre-differentiation culture and rooting culture sequentially. Compared with the existing organ culture, the invention has high propagation rate, rooting rate reaches 100%, root system quality is good, and differentiation rate of strong fleshy tap root is improved to 96%, thus ensuring survival rate after transplantation.

The invention belongs to the technical field of animal tissue isolation and culturing, and particularly relates to a method of porcine intestinal crypt isolation and 3D type organ culturing. In the course of porcine intestinal crypt isolation, penicillin and streptomycin are added, so that in the later-stage culturing course, pollution caused by intestinal microorganisms is avoided; rotating speedand time for treating an intestinal segment of a horizontal rotary instrument and frequency and time of a vortex of a vortex instrument are optimized, so that the obtained crypt is complete in shape,and higher in survival rate and bud rate. According to the method disclosed by the invention, a formula of a culture solution for culturing the crypt and the 3D type organs is optimized, so that thecrypt can be well polarized to form the 3D type organs, and the growth and the polarization are stable; the isolating condition and the optimal culturing method of the porcine intestinal crypt are determined, and favorable foundation for subsequently utilizing the porcine intestinal 3D type organs to study enterovirus and bacterial infection is established.

The invention provides a culture medium. The culture medium is prepared from a basic culture medium, a serum replacement, double antibodies, glutamate / ester, N-acetylcysteine, CHIR-99021 and LDN-193189. The culture medium provided by the embodiment has the advantages that the culture medium is used for in-vitro culturing of intestinal crypt or stem cells, so that the amount of the obtained intestineal crypts is greater than the amount of intestinal crypts in the prior art, the ratio of stem cells in the obtained intestinal crypts is higher, and the amount of the obtained intestinal stem cellsis more; the culture medium is used for the in-vitro culturing of the intestinal crypts, so that the dryness of the stem cells in the intestinal crypts can be maintained for a long time, the cell proliferation in the intestinal crypts is more vigorous, and the cell differentiation in intestinal crypts is normal and ordered; the culture medium is favorable for the popularization of an intestinal epithelial stem cell culture system and an intestinal crypt organ culture system, is favorable for the in-vitro massive proliferation of the intestinal stem cells, and provides a favorable platform forthe treatment of intestinal diseases.

The invention relates to the field of organoid culture, in particular to a culture medium, a method and a kit for rapidly culturing tumor organoids. According to the culture medium for culturing the tumor organoids provided by the invention, the time for culturing the tumor organoids can be greatly shortened. The kit for rapidly culturing the tumor organoid comprises a culture medium for culturing the tumor organoid, a tissue preservation solution, a tissue digestion solution and a cell mass collection solution, and when the kit is used for culturing the tumor organoid, the success rate and survival rate of culturing the tumor organoid can be increased, an organoid model can be rapidly established within 2-5 days, which can be applied to subsequent experimental tests.

The invention discloses a pleuroperitoneal fluid type organ culture medium and method and a drug sensitive testing method. The pleuroperitoneal fluid type organ culture medium can effective maintain specificity of tissue cells and characteristics of stem cells and ensure high consistency of genetic typing as well as high similarity of tissue shape. Besides meeting the demands of scientific research, the pleuroperitoneal fluid type organ culture medium can be applied to the aspect of clinical medication guidance, and through in vitro organ culture, provide patients with a beneficial option on medication guidance.

The present invention relates to cell and tissue culture. In particular, the present invention provides a method for preparing an organotypic culture using dissociated cells or microexplants obtained from an animal organ. The method for preparing an organotypic culture comprises culturing cells from an organ on a surface characterised in that the cells are compacted. The invention further relates to a high-throughput method for the preparation of a collection of organotypic cultures. The invention further relates to a device for carrying out a method of organotypic culture according to the invention.

The invention provides a culture medium for stomach cancer organs and a culture method. The culture medium comprises a basic culture medium 1640, specific addition factors and sterile water, wherein the mass ratio of the basic culture medium 1640 to the sterile water is 99:1; and the specific addition factors comprise B27 without vitamin A, N-acetylcysteine, EGFs (epidermal growth factors), Noggin, R-spondin 1, Wnt3a, CHIR99021, thiazovivin, Gastrin I, valproic acid, a penicillin and streptomycin mixed liquid, amphotericin B and Primocin. The culture medium can be adopted to culture the stomach cancer organs, morphology structures and gene characteristics of primary tissue can be maintained, the risk of microorganism pollution in stomach cancer culture can be effectively reduced, and the success rate and the survival rate of stomach cancer organ culture can be increased.

This invention discloses a method for extracting natural composite growth factor. The method comprises: constructing artificial tissue / organ in vitro through tissue-engineering method, collecting the supernatant during tissue / organ culture, separating and purifying to obtain natural composite growth factor. The method can obtain large quantities of amino acids or proteins including growth factor and nutrition factor, which, after separation and purification, can be used in cell culture, cosmetics, foods and drugs.

A curtain-type anchoring biological culturing reactor and its method for culturing vegetable cell, tissue or organs are disclosed. The reactor consists of bed(E), curtain-type netted double-layer anchoring culturing bed(A), supply system(B), hanger(C), composite circulation feeder(F) and disposable sterile sealing film(G). (A) is hung on (C) and has (E), (F) is arranged on top of (E), (E) is sleeved with (G). It's simple, safe and cheap. It has less consumption and can be used for continuous production.

The invention provides a method for culturing and subculturing intestinal cancer organoid derived from circulating tumor cells. The method comprises the following steps: (1) mixing a human colorectalcancer circulating tumor cell organoid culture medium with Matrigel according to a ratio of 1:1 to prepare a starting plate glue; (2) resuspending cells captured from the blood of a metastatic colorectal cancer patient by a cell filter with the pore diameter of 5.5 [mu]m by using the starting plate glue, and dropwise adding the cells into a 24-pore plate; (3) after the starting plate glue is solidified, adding the human colorectal cancer circulating tumor cell organoid culture medium around the starting plate glue, and culturing; (4) during subculturing, discarding the original culture medium,and adding a TrypLE digestive juice for digestion; and (5) stopping digestion by using a DMEM containing 10% of FBS, cleaning twice by using PBS, and re-suspending a starting plate by using the starting plate glue. The method for culturing and subculturing intestinal cancer organoid derived from the circulating tumor cells provides an experimental model for research on metastasis and drug resistance mechanisms caused by the circulating tumor cells, can perform drug sensitivity screening, and provides an effective basis for individualized treatment of advanced intestinal cancer.

The invention discloses an organoid culture chip and culture method. The organoid culture chip comprises a cell culture plate; and an organoid culture device arranged in the cell culture plate. A culture medium liquid storage tank is formed between the organoid culture device and the cell culture plate; and the organoid culture device comprises an organoid culture device body, an organoid culture chamber arranged in the organoid culture device body and side holes formed in side walls of the two sides of the organoid culture device body, the organoid culture chamber communicates with the side holes in the side walls of the two sides to form perfusion channels, the organoid culture chamber comprises a sample adding hole formed in the top and a micropore formed in the bottom, and both the sample adding hole and the micropore communicate with the bottom of the cell culture plate. According to the organoid culture chip and the culture method, high-throughput and one-step dynamic perfusion culture of organoids with a uniform form can be realized, and the whole growth and development process of the organoids can be dynamically observed in situ.

The invention discloses a culture method of an osteosarcoma organ and an osteosarcoma culture medium thereof. The invention uses two culture methods, namely a preparation method of a 3D culture modelof an osteosarcoma organ and a preparation method of a gas-liquid culture model of the osteosarcoma organ. The preparation method of the osteosarcoma organ 3D culture model comprises the following steps: (1) tissue cleaning: performing cleaning for 2 to 5 times by using 10ml of ADMEM / F12 containing 1 x Pen-Strep Glutamine; (2) cutting tissues into pieces with a size of 1mm<3>, performing digestionfor 30-60 minutes by using digestive juice, and performing centrifuging for 5 minutes at a temperature of 4 DEG C and a speed of 1200r after the digestion; (3) performing cleaning for three times byusing 10ml of ADMEM / F12 containing 1 x Pen-Strep Glutamine; (4) performing filtering with a 100 [mu]M cell sieve, performing counting and centrifuging, adding Collagen into 1-100 [mu] g / ml laminin, performing re-suspending, and paving a plate; (5) replacing the bone tumor culture medium once every 3-4 days in the culture period; and (6) performing passage on the organoids once every 15-20 days generally, and adding 2-5 times of TrpLE Express into each hole during passage.

The invention provides a culture and cryopreservation method for a prostatic cancer organoid with a tumor immune microenvironment. The culture and cryopreservation method comprises the following steps: separating mononuclear cells and autoserum from peripheral blood, and stimulating the mononuclear cells by using a CD28 antibody and IL-2; separating tumor cells from a prostatic cancer tissue sample; mixing the tumor cells with a culture medium and a temperature-sensitive hydrogel, and then carrying out culturing and subculturing to obtain a tumor organoid precursor; stimulating the tumor organoid precursor by using IFN-gamma, and then conducting dissociating to form stimulated tumor cells; mixing the stimulated tumor cells with the stimulated mononuclear cells, conducting stimulating witha PD-1 antibody, mixing the stimulated mixture with the temperature-sensitive hydrogel, and carrying out culturing; and cryopreserving the tumor organoid with a cryopreservation solution containing FBS and the temperature-sensitive hydrogel. With the method in the invention, the success rate of culturing of organoids with a tumor microenvironment can be greatly improved at low cost.

The invention relates to a method for preparing breast cancer bone metastasis organs, which comprises the following steps: mixing breast cancer bone metastasis cells, feeder layer cells, a temperature-sensitive hydrogel and a first culture medium, and culturing to obtain breast cancer bone metastasis organs; wherein the feeder layer cells comprise bone marrow mesenchymal stem cells without a proliferation function. By means of the technical scheme, the bone marrow mesenchymal stem cells without the proliferation function are used as feeder layer cells for breast cancer bone metastasis organ culture, and the success rate of breast cancer bone metastasis organ culture through the method is high.

The invention relates to a method for culturing a dedifferentiated or undifferentiated thyroid carcinoma organ. The method comprises the following steps: mixing dedifferentiated or undifferentiated thyroid carcinoma tissue cells with a thyroid carcinoma culture medium and matrix glue to obtain a to-be-cultured substance, wherein the thyroid carcinoma culture medium contains nicotinamide and BM-Cyclin, and based on the thyroid carcinoma culture medium, the concentration of nicotinamide is 5-20 mM, and the concentration of BM-Cyclin is 5-30 [mu]g / ml; and carrying out culture amplification on theto-be-cultured substance to obtain the dedifferentiated or undifferentiated thyroid carcinoma organ. In the method for culturing the dedifferentiated or undifferentiated thyroid carcinoma organ, theadopted thyroid carcinoma culture medium contains nicotinamide and BM-Cyclin with specific concentrations, so that the in-vitro growth of the dedifferentiated or undifferentiated thyroid carcinoma organ is promoted. Therefore, when the method provided by the invention is used for culturing the dedifferentiated or undifferentiated thyroid carcinoma organ, the culture success rate is relatively high.

The invention relates to a method for culturing liver cancer organoid. The method comprises the following steps: mixing liver cancer tissue cells with a culture medium and matrix gel to obtain a to-be-cultured substance, wherein the culture medium containing an insulin growth factor-2, and the concentration of the insulin growth factor 2 being 1-50ng / ml, preferably 5-10ng / ml based on the culture medium; and carrying out culture amplification on the to-be-cultured substance to obtain the liver cancer organoid. According to the culture method of the liver cancer organoid provided by the invention, the adopted culture medium contains the insulin growth factor-2 with the specific concentration, and the insulin growth factor-2 with the specific concentration can promotes in-vitro growth of theliver cancer organoid, so that the culture success rate is relatively high when the method provided by the invention is used for culturing the liver cancer organoid.