243 results about "Matrigel" patented technology

The invention relates to bionic matrix hydrogel for constructing bone organs and a preparation method of the bionic matrix hydrogel. The bionic matrix hydrogel is prepared from a bone acellular extracellular matrix, a salmon DNA (Deoxyribose Nucleic Acid) network and a calcium phosphate oligomer under the combination of photo-crosslinking and self-assembly physical crosslinking. The acellular extracellular matrix in the bionic matrix hydrogel can simulate organic components in natural bones, the calcium phosphate oligomer can simulate inorganic components in the natural bones, and sufficient phosphate ions can be provided for biomineralization due to introduction of a DNA network. The biomimetic matrix hydrogel can replace Matrigel to act in bone organoid culture, provide a living environment similar to bone tissue for three-dimensional culture cells, enhance substance exchange and signal transduction between cells, promote proliferation, adhesion and osteogenic differentiation of bone marrow mesenchymal stem cells, and further accelerate the process of biomineralization. The invention provides an optional new material for preparing the functional bone organ.

The invention relates to an organ-like core-shell microsphere as well as a preparation method and application thereof. The preparation method comprises the steps that 1, raw materials are prepared, specifically, shell raw materials and inner core raw materials are prepared, the shell raw materials comprise a photoinitiator and methacrylated hyaluronic acid (HAMA), and the inner core raw materials comprise organoid precursor cell suspension and matrigel; 2) molding: extruding the dispersion phase of the shell raw material wrapping the core raw material to a continuous phase by using a micro-fluidic chip and a high-precision injection pump, cutting the dispersion phase into liquid drops by the continuous phase, and performing illumination curing on the liquid drops to form core-shell microspheres; and (3) incubating, namely culturing the core-shell microspheres in a culture medium, so that the organoid precursor cells in the inner core are developed into organoids. According to the preparation method, rapid forming and curing of the organ-like precursor cells carried by the matrigel are achieved, so that the time of the organ-like precursor cells staying in the oil phase is shortened, the cells can obtain oxygen and nutrient substances, and the cell activity is improved.

The invention relates to the field of cell biology, and particularly discloses a method for preparing mesoderm progenitor cells from human pluripotent stem cells and an induced differentiation culture medium of the mesoderm progenitor cells. The method comprises the following steps: inoculating human pluripotent stem cells in a single cell state into a culture dish covered with matrigel; and inducing the human pluripotent stem cells to be differentiated into the mesoderm progenitor cells within 1-3 days by using the mesoderm progenitor cell induced differentiation culture medium. Wherein the mesoderm progenitor cell induced differentiation culture medium is composed of a basic culture medium and an additive, and the additive contains a B-27 additive, non-essential amino acid, glutamine, ascorbic acid, polyvinyl alcohol, a GSK-3 inhibitor and a CDK8 inhibitor. According to the method, the human pluripotent stem cells are subjected to induced differentiation by using the mesoderm progenitor cell induced differentiation culture medium with specific composition, the method does not depend on cloning of the pluripotent stem cells, a large number of high-uniformity mesoderm progenitor cells can be obtained through induced differentiation within 1-3 days, the induction efficiency is high, and the stability is high.

The invention belongs to the technical field of biological medicine, and particularly discloses an artificial valve tissue calcification model and a preparation method and application thereof. The preparation method comprises the following steps: providing human valvular interstitial cells; the method comprises the following steps: providing a hydrogel system, wherein the hydrogel system comprises type I collagen, fibrinogen and Matrigel matrigel; constructing an engineered heart valve tissue strip by taking the hydrogel system as a skeleton structure between valve interstitial cells; and culturing the engineered heart valve tissue strip by using a calcification induction culture solution, so that the valve tissue generates a calcification phenotype. The model provided by the invention can generate a valve calcification phenotype which is similar to that after human heart valve calcification and is represented by valve interstitial cell activation, extracellular matrix remodeling such as collagen mass secretion and calcium deposition region generation. The model provided by the invention is low in cost, can be produced in batches, meets the research on the pathogenesis of extracorporeal valve calcification, and makes up for the dilemma that no artificial valve calcification model is available for drug research and development at present.

The invention discloses a construction method of a homogenized self-assembled heart organoid, and belongs to the field of organoid. The preparation scheme of the heart organoid provided by the invention is controllable in method and simple in procedure, emphasizes the transition of differentiation from an iPSC 2D level to a 3D self-assembly heart organoid, overcomes the heterogeneity, including size, structure, function and even gene difference, of cross-batch or even same-batch organoid caused by an existing differentiation method, and has good cross-batch reproducibility. The constructed heart organoid contains various cell types and can continuously beat for at least 180 days in vitro, and the heart organoid in the same batch is uniform in height and stable in function. The preparation scheme does not need an engineering scaffold, matrigel and the like, emphasizes self-assembly of heart organs, and better accords with physiological characteristics of a human body.

The invention provides a micro-fluidic chip capable of culturing oral squamous cell carcinoma organoid and a preparation method thereof, and relates to the technical field of micro-fluidic chips, the chip comprises a four-layer structure: the first layer is a closed layer, the second layer is a culture solution / drug pool layer (containing a concentration gradient injection channel and a slow flow channel), the third layer is a matrix culture pool layer, and the fourth layer is a matrix culture pool layer. The fourth layer is a matrigel inflow layer (containing an S-shaped flow channel); the second layer and the third layer are integrally formed, and all the layers are connected through oxygen plasma welding; the problems that in the prior art, matrigel is prone to shifting, drug gradient regulation and control is poor, operation is tedious, and adaptability is insufficient are solved, the matrigel and organoids can be stably fixed, the drug concentration gradient can be precisely regulated and controlled, the nutrition supply efficiency is enhanced, the structural integration level and operation convenience are improved, and the application prospect is wide. The method is suitable for oral squamous cell carcinoma organoid culture, drug screening and tumor mechanism research.

The invention provides preparation, sorting and application of LGE / CGE organoid and GABAergic neurons, and belongs to the field of brain organoid differentiation and GABAergic neurons. The preparation method of the LGE / CGE type organ comprises the following steps: S1, performing embryoid formation and initial culture on the 0th-4th day; s2, performing nerve induction and matrigel embedding on the 5th-11th day; s3, culturing the brain organoid on the 12th day to the 19th day; s4, dynamically culturing and screening the brain organoid on the 20th-35th day; and S5, carrying out differentiation and collection on LCOs of the LGE / CGE type organs after the 35th day. According to the method, the LGE / CGE organs containing the high-purity GABA energy neurons are obtained through differentiation from the brain organs for the first time, the high-purity LGE type GABA energy neurons and the high-purity CGE type GABA energy neurons are obtained through a specific sorting method, and the method has the deep application potential in the fields of disease model construction, medicine research and development and the like.

The invention belongs to the technical field of analysis and detection, and particularly relates to a beta-galactosidase response type hemicyanine near-infrared fluorescent probe, a preparation method thereof and application of the beta-galactosidase response type hemicyanine near-infrared fluorescent probe as a tumor organ imaging probe. The near-infrared fluorescent probe provided by the invention takes a water-soluble hemicyanine skeleton as a fluorophore, so that the biocompatibility of the near-infrared fluorescent probe is improved; based on the principle that beta-galactose can be selectively cut by beta-galactosidase, a galactose structure with a self-leaving group is used as a response group, selective quick response to the beta-galactosidase is achieved, and high sensitivity to the beta-galactosidase is shown. The near-infrared fluorescent probe provided by the invention can emit near-infrared fluorescence with long wavelength, has lower background fluorescence and good penetration depth, and can be directly used for detecting organoids containing the matrigel without eliminating the matrigel, so that the organoids are prevented from being damaged or additional experimental variables are prevented from being introduced in the process of eliminating the matrigel.

The invention relates to a culture method of a pancreatic cancer organ model. The culture method comprises the following steps: cleaning tissues and chopping the tissues through a chopping device; transferring the chopped tissue into a centrifugal tube filled with tissue digestive juice for digestion, and collecting upper suspension and residual tissue precipitate; mixing the collected upper-layer suspension with the digested residual precipitate, and treating to obtain cell and tissue precipitates; embedding the tissues and the cells into matrigel; and after the matrigel is solidified, culturing tissue cells by using a corresponding organoid culture medium, and obtaining primary organoid tissues which are pancreatic cancer tissues. Due to the adoption of the technical scheme, the culture method of the pancreatic cancer organoid model can be used for stably culturing pancreatic cancer organoid with high success rate, and the invention also relates to a culture method of the pancreatic cancer para-carcinoma organoid model.

The invention belongs to the technical field of cell culture, and particularly relates to a method for promoting proliferation and differentiation of cervical squamous epithelial cells. The method comprises the following steps: S1, extracting and culturing primary cervix uteri squamous epithelial cells from cervix uteri tissues to obtain primary cervix uteri squamous epithelial cells; s2, adding the primary cervical squamous epithelial cells into a cell proliferation induction culture solution to induce cell proliferation, so as to obtain proliferated cervical squamous epithelial cells; and S3, uniformly mixing the proliferated cervical squamous epithelial cells with differentiation matrigel, and inducing cell differentiation by using a cell differentiation induction culture solution to obtain differentiated cervical squamous epithelial cells. Proliferation and differentiation of the cervical squamous epithelial cells are promoted by preparing a proper culture medium, and influence factors of proliferation and differentiation of the cervical squamous epithelial cells are researched by culturing the cervical squamous epithelial cells in vitro, so that the pathogenesis of cervical erosion and cervical cancer can be clarified; therefore, a direction is provided for research and development of medicines for treating cervical erosion and cervical cancer.

The invention belongs to the technical field of cross culture of biological materials and organoids, and particularly relates to an ECM protein for three-dimensional culture of vascular organoids. HA-; the invention relates to a preparation method and application of Gel composite hydrogel. In order to solve the problems that in the 3D culture process of existing cell spheres or organoid spheres, matrigel derived from mouse sarcoma seriously depends on, components are undefined, the price is high, and immunogenicity exists, the composite gel with the definite components is provided and used for replacing Matrigel to conduct 3D culture of vascular organoids. The composite hydrogel prepared by the invention has the characteristics of short gelation time, good biocompatibility, adjustable gel components and the like under the synergistic effect of all the raw materials, can solve the problem of matrigel used for 3D culture of organoids, can replace the potential of matrigel in the future, reduces the price of 3D culture cell scaffold materials, and has broad application prospects. Meanwhile, the method can also be used in the regenerative medicine field of other two-dimensional or three-dimensional cell and organoid culture.

Provided are chemically modified silk fibroin and a use thereof in cell and organoid culture. The chemically modified silk fibroin is a product obtained by sequentially carboxylating silk fibroin and modifying same by an organic amine having a phenol group. Also provided is a hydrogel based on the chemically modified silk fibroin. A hydrogel system has a hierarchical structure and mechanical properties similar to those of an extracellular matrix, and exhibits characteristics such as definite composition, controllable physicochemical properties, low cytotoxicity, good biocompatibility, and biodegradability, can support the growth and differentiation of cells and organoids, and is suitable as a matrigel for cell and organoid culture.

The invention discloses a construction method of a trophoblast organ-like model, the trophoblast organ-like model and application thereof, and relates to the technical field of cell model.The construction method comprises the steps that placenta villus tissue is aseptically separated, gradient separation and purification are conducted to obtain extraembryonic early cells, the extraembryonic early cells are mixed with matrigel, and the trophoblast organ-like model is obtained; and carrying out 3D culture by using a culture medium containing early-stage and later-stage differentiation compositions to obtain the organ-like model with various trophoblast cell lineages. Based on the model, placenta toxicity research is carried out on pregnancy drugs, and the research specifically comprises the degree of 3D organoid form destruction after drug treatment, organoid activity and the expression condition of trophoblast marker genes. Compared with a traditional two-dimensional cell model, the placental toxicity detection method has the advantages that the placental toxicity of the medicine is detected by taking the trophoblast-like organ as the model, the accuracy is higher, the predictability is higher, and the influence of the medicine on human placental tissues is accurately reflected.

The invention provides a mouse middle ear epithelium 3D organoid and a construction method thereof, and is characterized in that the method comprises the following steps: a, taking a bubble structure of a mouse middle ear tissue, cleaning, digesting, filtering, centrifuging filtrate, collecting precipitate, and separating primary cells of the mouse middle ear; and b, inoculating the digested single cells and cell agglomerates into a 24-pore plate, wrapping with matrigel, and carrying out 3D culture, wherein a medium is a middle ear epithelium amplification medium. And culturing for about 3-5 days to obtain the mouse 3D middle ear proliferation type organoid. And performing differentiation culture by using a middle ear epithelium differentiation medium to obtain the mouse middle ear epithelium organ. According to the method, the 3D mouse middle ear epithelium organ is successfully constructed, the physiological and pathological processes of the middle ear are conveniently analyzed and researched, the interaction between the middle ear microenvironment and cells can be more accurately simulated, and the application range is wide as a model.

The invention discloses a culture system, culture method and application of gastrointestinal tumor chemotherapy drug-resistant organoid, the culture system comprises: a basic culture medium, which is a DMEM / F12 culture medium, and is additionally added with a special growth promoting factor obtained from a natural extract; the multiple growth factors comprise an epidermal growth factor (EGF), a fibroblast growth factor (FGF), a bone morphogenetic protein (BMP) and a vascular endothelial growth factor receptor inhibitor (VEGFRi), and the concentration of each growth factor is accurately screened and optimized; the extracellular matrix is Matrigel modified by physical and chemical methods; the specific additive comprises an antioxidant N-acetylcysteine (NAC). According to the culture system, the culture method and the application of the gastrointestinal tumor chemotherapy drug-resistant organoid, the culture system which is closer to a real environment in vivo is constructed by optimizing basic culture medium components, setting various growth factors and specific additive concentrations and adopting modified Matrigel as an extracellular matrix.

The invention discloses a preparation method and application of a bovine tongue mucous membrane organoid, and relates to the technical field of organoid separation and culture. The preparation method of the ox tongue mucosa organoid comprises the following steps: treating fetal ox tongue tip tissues with digestive enzymes Dispase I, Collagenase II and Collagenase IV, separating ox tongue mucosa, cutting into pieces, treating again with pancreatin, and sieving to remove tissues and impurities; and continuously culturing by adopting a three-dimensional culture technology-matrigel embedding method and an organoid culture medium to obtain the final ox tongue mucous membrane organoid. The ox tongue mucous membrane organoid prepared by the preparation method disclosed by the invention has normal cell proliferation, differentiation and metabolism functions, and can be stably passaged under an in-vitro culture condition; the method can be widely applied to various related scientific researches such as FMDV infection mechanism research, drug screening and vaccine evaluation.

The invention relates to the technical field of biology, and particularly discloses a construction method and application of an endometrial epithelium organ fibrosis model. According to the method, human endometrial epithelial cells are extracted, and an endometrial epithelial organ is established by adopting a Matrigel three-dimensional culture system. Afterwards, the organoid is induced to form endometrial phenotypes in different physiological cycles through hormone stimulation, and a ferroptosis inducer FIN56 is added in the culture process to induce fibrosis. The expression of GPX4, Fibronectin and p-Smad2 / 3 is detected through Western blot and immunofluorescence, and the successful construction of the fibrosis model is verified. The invention provides an in-vitro model capable of simulating endometrial fibrosis pathological characteristics of a patient with PCOS (polycystic ovarian syndrome), the in-vitro model can be used for mechanism research and drug screening of PCOS-related endometrial lesions, and a new research tool and an intervention strategy are provided for treatment of PCOS endometrial fibrosis.

The invention relates to the technical field of organs, and discloses construction of a matrigel-free kidney organoid and a method for evaluating drug safety by using liver and gall-kidney organoid. By utilizing the culture medium set provided by the invention, the kidney organoid with more complete functions and structures can be constructed through iPSC induction, and matrigel does not need to be adopted in the construction and culture process, so that the construction operation of the organoid is greatly simplified, and the cost is saved. The kidney-like organ constructed by the invention has a basic structure of the kidney, has corresponding response to medicine components which are known in the field and have influence on the kidney, and can be used as a kidney in-vitro model.

The invention discloses a bladder cancer lymph node metastasis derived tumor organoid culture medium, a culture method and a rapid drug detection method, and belongs to the technical field of organoid and cell culture. The method comprises the following steps: culturing a bladder cancer lymph node metastasis-derived tumor organoid by using a bladder cancer lymph node metastasis-derived tumor organoid culture medium, immersing matrigel drops in the culture medium, and obtaining the bladder cancer lymph node metastasis-derived tumor organoid after the cultured organoid grows to 100-150 microns, to-be-detected drugs with different concentrations are directly added into the cell pore plate inoculated with the tumor organoids, the influence of the to-be-detected drugs on cell growth or metabolism is analyzed through a full-view bright-field imaging method, and the detection result of the drug for treating bladder cancer lymph node metastasis is obtained. Compared with a traditional ENR organ-like culture medium, the method has the advantages that the effect is better, and the cost is lower; the kit is used for screening medicines for bladder cancer lymph node metastasis, is simple and efficient, and can be used for accurately detecting medicines for efficiently killing bladder cancer lymph node metastasis cells.

The invention relates to the technical field of biotechnology or biomedical engineering, in particular to hydrogel, bio-ink, organoids constructed by using the hydrogel, a drug screening model and a construction method. The hydrogel is prepared from 3 to 4 weight percent of gelatin, 0.5 to 2 weight percent of sodium alginate, 20 to 40 weight percent of matrigel and a cell culture medium. The hydrogel is helpful for improving the bionic degree of a tumor microenvironment, can be used for preparing bio-ink containing target cells such as tumor cells, particularly can be used for preparing bio-ink of primary cells (such as primary renal cell carcinoma cells), and an organ-like model constructed by using the bio-ink under a proper 3D biological printing condition is uniform in growth and high in biocompatibility. The cell growth speed is high, the consistency with parent tissues is high, the in-vitro amplification time of the patient-derived tumor cells can be shortened, the influence of subculture factors on the cell affinity can be reduced, the drug development progress can be accelerated, and the production cost can be reduced.

The present invention discloses a method for culturing vitreoretinal organoids in vitro, comprising the following steps: (1) amplifying and culturing pluripotent stem cells to a cell confluence of 50% to 90%; (2) on day 0, digesting the stem cells with a digestion solution; adding a neurosphere culture medium for culturing; adding matrigel for culturing, and then adding a neurosphere culture medium for culturing to obtain a cell suspension; (3) adding a ROCK pathway inhibitor to the cell suspension, and culturing in the neurosphere culture medium for 3 to 5 days to obtain ectoderm neurospheres; (4) replacing the neurosphere culture medium with a vitreous culture medium I, adding TGFB1 protein, and placing the cell suspension on a shaker for 25 to 28 days; (5) replacing the vitreous culture medium I with a vitreous culture medium II, and culturing the cell suspension for 60 to 300 days to obtain vitreoretinal organoids. The vitreous organoids of the retina cultured in vitro according to the present invention can simulate the vitreous in vivo.

The invention relates to a lung cancer organoid and peripheral blood source immune cell co-culture model and a construction method thereof. Specifically, the invention provides a construction method of a macrophage and tumor organoid co-culture model for evaluating anti-tumor activity, monocytes of autologous or allogeneic peripheral blood of a patient are induced and differentiated into high-purity macrophages in vitro, and the high-purity macrophages and lung cancer tumor organoid are subjected to three-dimensional co-culture in matrigel; the model aims at highly reducing a core interaction network of tumor cells and macrophages in TME; the dynamic change of the polarization state of the macrophage is simulated and observed; the problem that an organ-like model is incomplete due to immune component deficiency or spatial positioning distortion of an existing model is effectively solved, an experimental platform closer to the physiological state is provided for tumor immune microenvironment research, and therefore development of the tumor immune treatment field is promoted.

The invention discloses a construction method of a human microglial cell model carrying ENTPD1 gene mutation, and the human microglial cell model is prepared by the following steps: extracting PBMC from venous blood of a hereditary spasmodic paraplegia patient, then electrically transforming four factors of Oct4, Sox2, Klf4 and cMyc, and culturing in a Matrigel-coated culture dish for 15-20 days to obtain induced pluripotent stem cells; the induced pluripotent stem cells can form a unique human microglial cell model, and the human microglial cell model and the induced pluripotent stem cells are co-cultured to obtain an organoid, so that the analysis of a pathogenic molecular mechanism behind a disease risk gene is greatly facilitated, the defects of a rodent-derived microglial cell model are complemented, and the development of the rodent-derived microglial cell model is promoted. And the influence of the ENTPD1 on the related functions of the microglial cells is further discussed.

The invention discloses a carbon dot, hydrogel containing the carbon dot as well as a preparation method and application of the hydrogel, and relates to the technical field of carbon materials. The carbon dot is prepared from the following raw materials: a hydroxyl-containing flavonoid compound and epsilon-polylysine; the carbon dots contain aniline bonds formed by hydroxyl groups of the hydroxyl-containing flavonoid compounds and amino groups of the epsilon-polylysine. The carbon dot shows higher tumor cell killing property and lower normal cell toxicity than the raw material hydroxyl-containing flavonoid compound at the cellular level, can obviously induce immunogenic death of cancer cells and does not induce up-regulation of extracellular matrix collagen, and also has a better effect of regulating a tumor immune microenvironment.

The invention discloses a construction method and application of an animal model for evaluating cardiotoxicity of antitumor drugs, sodium carboxymethyl cellulose (CMC-Na) with extremely low cost is adopted to replace matrigel, a tumor-bearing mouse model with high stability can be constructed at low cost, and experimental verification shows that 4T1 cells are resuspended by using a 0.7% CMC-Na solution, and the tumor formation rate of subcutaneous inoculation reaches gt; 95%. The tumor microenvironment-mediated cardiac injury critical period is accurately captured, and the constructed tumor-bearing mouse model is the optimal tumor-bearing mouse model for evaluating the cardiotoxicity of an antitumor drug (adriamycin) in the tumor microenvironment in the third week after the mouse has a 50mm < 3 > tumor.

The invention discloses a preparation method of a mouse skin organoid, which comprises the following steps: separating dermis and epidermis of an E18.5 mouse to obtain dermis cells, mixing the dermis cells with back skin cells of an E14.5 embryo mouse, mixing by using Matrigel to form cell spheres, and gradually forming the skin organoid in in-vitro culture; the organoid is mainly applied to batch production of organoid containing hair follicles. The invention relates to the technical field of stem cells and regenerative medicine. According to the preparation method of the mouse skin organoid, a cell source is improved in the experiment process, the mouse skin organoid containing hair follicles can still be formed after cells in the early development stage are added, and related developmental biology research is carried out. And the source of another dermal cell is not limited to late pregnancy, and also can be a newborn mouse, so that the source of seed cells is expanded, and the cost is reduced.

The invention relates to a co-culture microsphere model with a core-shell structure and a preparation method of the co-culture microsphere model. The co-culture microsphere model comprises a core-shell microsphere, the core-shell microsphere is composed of an inner core and an outer shell, the inner core is an organoid loaded by matrigel, the outer shell is formed by crosslinking a photo-crosslinking hydrogel material, co-culture cells are doped in the photo-crosslinking hydrogel material, and the inner core is wrapped by the outer shell. According to the co-culture microsphere model, double-layer structure microspheres of co-culture cell wrapped organoids are formed, immune infiltration and in-vivo characteristics can be simulated, and a similar in-vivo three-dimensional tumor immune microenvironment is provided. And the types of cells loaded on the inner layer and the outer layer can be flexibly replaced to form a microsphere model for co-culture of single or multiple cells and organoids. The co-culture microsphere model is more stable in structure, the size of the diameter of the core-shell microsphere can be optimized by changing 3D printing parameters, the operability of subsequent research is high, and the co-culture microsphere model is suitable for different research application scenes.

The invention discloses a tumor ultrasonic image sensitivity detection method, and relates to the technical field of tumor ultrasonic imaging. Comprising the following steps: S1, preparing tumor balls for simulating tumors, and forming the tumor balls in a complete culture medium containing matrigel by using a 4T1 breast cancer cell strain through a suspension culture method; according to the method, 4T1 breast cancer cell tumor spheres with different sizes are accurately prepared, the sizes of nano and micron contrast agents are detected by utilizing a dynamic light scattering technology, the accuracy and reliability of an experiment are ensured, the penetration condition of a contrast material in the tumor spheres can be intuitively evaluated through penetration detection of a laser confocal microscope, and the accuracy and reliability of the experiment are ensured. Through combination of ultrasonic imaging and ultrasonic quantification software, the gray value of the region of interest is accurately quantified, and accurate data is provided for subsequent statistical analysis; in addition, unidirectional variance analysis is carried out through Graphpad software, the sensitivity difference of ultrasonic images under different conditions can be scientifically evaluated, a powerful basis is provided for early diagnosis and treatment of tumors, and the method has a wide application prospect.

The invention discloses a composition and a method for constructing ovarian organoid based on a clinical tissue sample. The composition comprises a sample preservation solution, a sample digestive juice and a sample culture medium, the method comprises the following operation steps: transporting a preservation solution A to a laboratory, and pretreating to obtain tissue fragment precipitates; carrying out resuspension incubation on the tissue fragment precipitate by using digestive juice B for digestion; collecting tissue blocks which are not completely digested and filtered, and performing secondary digestion to obtain single cells; combining the cells obtained in two times to obtain the required single cell; adding matrigel, then adding the mixture into a pore plate, and putting the pore plate into an incubator until the matrigel is solidified; and taking out the pore plate, adding a culture medium L2, and culturing to obtain the ovarian organ constructed based on the clinical sample. According to the method, the ovarian organoid is efficiently constructed from the ovarian tissue sample; the provided culture solution formula makes up the dilemma that normal ovarian tissue organoids are difficult to form and normal tissue-derived organoids are difficult to passage at present, and reduces the burden and experiment cost brought by sampling in the disease research process.

The invention belongs to the technical field of bioengineering, and relates to heparan sulfate defect type matrigel as well as a preparation method and application thereof. The preparation method comprises the following steps: providing natural matrigel; providing immobilized heparinase, wherein the immobilized heparinase can specifically catalyze and degrade heparan sulfate in the natural matrigel; carrying out contact reaction on the natural matrigel and the immobilized heparinase; and after the reaction is finished, separating the immobilized heparinase. According to the preparation method, the characteristics that the immobilized enzyme is convenient to separate and recycle are utilized, the problems that in a traditional solution enzyme method, enzyme residues exist, the reaction is difficult to accurately control, and subsequent purification steps are complex are solved, and a standardized heparan sulfate defective matrigel product which is controllable in component, free of residual enzyme and high in stability is provided. The method has wide application value in the fields of tumor mechanism research, anti-tumor drug screening, tissue engineering and the like.