108 results about "Adherent Culture" patented technology

The invention relates to a heart organoid and an in-vitro preparation method thereof, and belongs to the technical field of stem cells. The invention provides a heart organoid. The heart organoid is obtained by induced culture of an adherent pluripotent stem cell mass; the adherent pluripotent stem cell mass is obtained by adherent culture of a suspended pluripotent stem cell mass; the suspension pluripotent stem cell mass is obtained by suspension culture of pluripotent stem cells. According to the in-vitro preparation process of the heart organoid, after pluripotent stem cells form balls, the pluripotent stem cell balls are subjected to staged induced differentiation under adherent culture, so that cell subtypes such as epicardial cells and the like can be generated without additional addition or additional stimulation of signal channels, and the heart organoid is more homogeneous. The self-occurring multi-lineage coexisting heart organoid can better simulate biological events derived from multiple cell types and multiple cell subtypes in the early development process of an embryonic heart, and has great application prospects in exploration of pathogenic factors and potential treatment targets of congenital heart diseases.

The invention relates to a method for rapidly detecting the cell adhesiveness of probiotics and application of the method. The method comprises the following steps: adding the probiotics into adherent culture cells, incubating, discarding liquid, digesting and collecting the cells by pancreatin, extracting total DNA (Deoxyribonucleic Acid), performing qPCR (Quantitative Polymerase Chain Reaction) amplification, and calculating the adhesion quantity of the probiotics on a single cell according to a Ct value. The invention provides a method for rapidly detecting the cell adhesion rate of probiotics by using a qPCR (quantitative polymerase chain reaction) technology, which comprises the following steps of: extracting total genome DNA (deoxyribonucleic acid) after the probiotics are adhered to cells on the basis of the gene expression difference of prokaryotic and eukaryotic cells, and detecting the specific gene expression level of the probiotics and the cells by using the qPCR technology. The cell adhesion ability of the probiotics is evaluated by calculating the ratio of the gene expression quantities of the two, and the method has the characteristics of high efficiency, rapidness, high accuracy and the like.

The present invention discloses a complete medium for culturing DF-1 cells, including DMEM basal medium 13.7g / L, insulin, non-essential amino acids, transferrin, glutathione, ascorbyl phosphate, sodium selenite, ethanolamine, hydrocortisone, and newborn calf serum. Wherein non-essential amino acids include glycine, alanine, proline, serine, and asparagine. A complete medium for culturing DF-1 cells provided by the present invention can well promote the proliferation of DF-1 cells, can achieve the same proliferation effect as high-concentration fetal bovine serum, and is suitable for the adherent culture of DF-1 cells; using low-concentration newborn calf serum, it is possible to greatly avoid the batch differences, unstable culture results, cytotoxicity, and high risk of carrying pathogens brought by the culture medium with high-concentration serum, narrow application range, expensive fetal bovine serum, and high culture cost; the components are simple, the culture medium preparation process is short in time consumption, the culture efficiency is high, the repeatability is good, and it is suitable for batch production.

The invention discloses a method for differentiating induced pluripotent stem cells into megakaryocytes and platelets, and relates to the technical field of induced pluripotent stem cell directional differentiation, and the method comprises the following steps: inoculating an iPSC single cell into a culture vessel, and carrying out adherent culture through a culture solution; after the cell adheres to the wall and before cloning is not formed, the liquid is changed into a first induction culture liquid, and the cell is induced and differentiated into a mesoderm cell; changing the solution into a second induction culture solution to induce and differentiate the mesoderm cells into hematopoietic endothelial cells; collecting the suspension cells, culturing the suspension cells in an amplification culture solution, and enriching megakaryocytes; inducing megakaryocyte polyploidy through a third induction culture solution; and culturing the polyploidy megakaryocyte through a fourth induction culture solution to promote platelet release. The method for inducing the pluripotent stem cells to differentiate into megakaryocytes and platelets is convenient and rapid, the megakaryocytes and platelets can be obtained through iPSC in a short time, and feeder cells, serum and additional gene editing are not needed; meanwhile, the yield is high.

The invention belongs to the technical field of cell biology, and particularly discloses a culture medium and a culture method of fish primary liver cells. The culture medium of the fish primary liver cells comprises the following components: 90% of an M199 culture medium, 10% of fetal calf serum, 100 U / mL of penicillin, 0.1 mg / mL of streptomycin, 5 ng / mL of EGF, 10 [mu] U / ml of insulin and 10 mM of dexamethasone. Based on the culture medium, an efficient and stable fish liver cell culture method is developed by combining a standardized anesthesia and disinfection process, a precise dissection and liver sampling method, an optimized tissue digestion and cell separation scheme, an efficient cell purification and adherent culture system, a stable passage technology and the like; a standardized hepatocyte line establishment operation process is defined, and the problems that species binding exists in an existing fish liver cell culture method, and a standardized culture process suitable for multiple fish fingerlings is difficult to form are solved.

The invention discloses a tissue preservation solution in organoid construction as well as a preparation method and application thereof. The tissue preservation solution is prepared from the following components: an Advanced DMEM / F12 culture medium, a mycillin / amphotericin B three-antibody mother solution, Glutamax-1, an HEPES buffer solution and a Y-27632 mother solution. The preparation method of the tissue preserving fluid comprises the following steps: adding an Advanced DMEM / F12 culture medium into a centrifugal tube on a super clean bench; then sequentially adding a mycillin / amphotericin B three-antibody mother solution, Glutamax-1, an HEPES buffer solution and Y-27632, and uniformly mixing, so as to obtain the tissue preservation solution. After uniformly mixing, filtering and sterilizing by using a disposable needle type filter, and collecting the preserving fluid into a centrifugal tube; the invention also discloses an application of the tissue preserving fluid in organoid construction. According to the tissue preserving fluid obtained by the invention, in the transportation process, the cell viability in the tissue is improved, the cell damage is reduced, and the success rate of primary cell adherent culture and organoid construction is increased.

The invention provides a method for inducing human pluripotent stem cells to be differentiated into retinal pigment epithelial cells and application. The method comprises the following steps: performing adherent culture on pluripotent stem cells to serve as differentiation starting cells; the differentiation starting cells are cultured in stages with different culture media to obtain the retinal pigment epithelial cells, and the culture media in at least one stage comprise a WNT non-classical pathway activator. The invention further provides a composition and a culture medium for inducing the human pluripotent stem cells to be differentiated into the retinal pigment epithelial cells, the differentiation efficiency of the RPE can be remarkably improved, the cell harvesting purity is improved, and the composition and the culture medium are suitable for large-scale industrial production.

The invention discloses a preparation method and application of mesenchymal stem cells based on neural crest organics.The preparation method comprises the following steps that 1, human induced pluripotent stem cells are added into digestive juice, and after digestion, a culture medium is added for dilution; 2, collecting cells, centrifugally separating the cells, removing supernatant, suspending the cells in a culture medium, and adjusting the size of a cell block to 50-80 microns; 3, culturing the cells, and then carrying out induced differentiation on neural crest organs; 4, digesting the neural crest organs into single cells, transferring the single cells into a serum-free mesenchymal stem cell culture medium for adherent culture, and then performing amplification to obtain the mesenchymal stem cells. According to the invention, the induced pluripotent stem cells ihPSCs3D are induced and differentiated to obtain neural crest organs, the neural crest-derived mesenchymal stem cells are obtained, and the obtained NCC-MSCs have improvement and repair effects on pancreatitis.

The present invention provides a method of producing a population of mesenchymal stromal cells (MSC) of CD73 + CD44 +, CD90 +. The CD73 + CD44 +, CD90 + MSC is used in a method for producing terminally differentiated osteoblasts, adipoblasts and chondroblasts from pluripotent stem cells (PSCs). The differentiation methods include the use of a single agent-a WNT signaling pathway activator used on an adherent culture of PSC, such as a GSK3 [beta] inhibitor.

The present invention relates to CD73 + CD44 + , CD90 + A method for generating a population of mesenchymal stromal cells (MSCs) is provided. + CD44 + , CD90 + MSCs are used in methods to generate terminally differentiated osteogenic, adipogenic, and chondrogenic cells from pluripotent stem cells (PSCs), which involve the use of a single agent, a WNT signaling pathway activator (e.g., a GSK3β inhibitor), in adherent cultures of PSCs.

The invention discloses a construction method and application of a human lung 3D cell microsphere and a human pulmonary fibrosis 3D organoid. The construction of the human lung 3D cell microspheres comprises the following steps: respectively digesting human primary fibrosis cells and human II-type alveolar epithelial cells which are subjected to adherent culture into single cells; mixing digested human primary fibrotic cells, human II-type alveolar epithelial cells and THP-1 macrophages subjected to suspension culture according to a cell number ratio of 1: 1: 1, so as to obtain a cell precipitate; and preparing the cell precipitate into cell matrigel microspheres by using a T-type microfluidic technology, and culturing to obtain the human lung 3D cell microspheres. And further inducing the human lung 3D cell microspheres by using a human lung organoid culture medium containing 50ng / mL of TGF-beta for 24-48 hours to obtain the human pulmonary fibrosis 3D organoid. According to the invention, 3D cell microspheres with human normal lung cell characteristics and protein expression can be formed within 3 days, and pulmonary fibrosis 3D organs can be obtained after short-time induction.

This invention relates to the interdisciplinary field of cell biology, experimental oncology, and translational medicine, specifically to a cancer-associated fibroblast cell line, its construction method, and its applications. This invention fills the research gap in mouse-derived oral squamous cell carcinoma-associated fibroblast cell line models and provides a method for constructing such a cell line. Through the inverted adherent culture and enzymatic digestion methods of this invention, a mouse-derived oral squamous cell carcinoma-associated fibroblast cell line with a clear origin, high purity, and long-term phenotypic and functional stability can be obtained. This allows for the establishment of a cryopreservable and resuscitable mouse oral squamous cell carcinoma-associated fibroblast cell bank, providing technical support for research related to the tumor microenvironment and immunotherapy.

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.

The invention relates to the technical field of adipose-derived mesenchymal stem cells, in particular to a method for efficiently enriching Sca-1 positive adipose-derived mesenchymal stem cells. The method comprises the following steps: selecting proper animal mice; carrying out primary ADSCs separation on the mouse; the method comprises the following steps: performing adherent culture on stromal vascular components, and separating Sca-1 + ADSC through a magnetic bead sorting method to form ADSC-AM cells; the positive rate in the ADSC-AM is detected through flow cytometry; and detecting the proliferative activity, osteogenic differentiation, adipogenic differentiation and chondrogenic differentiation capabilities in the ADSC-AM cells. According to the method for efficiently enriching Sca-1 positive adipose-derived mesenchymal stem cells provided by the invention, it is innovatively found that a cell population enriched by using Sca-1 as a marker of ADSC (adipose-derived mesenchymal stem cells) has better adipose-derived stem cell characteristics, and the cell population has the advantages of being capable of efficiently enriching Sca-1 positive adipose-derived mesenchymal stem cells, high in specificity, high in specificity, high in specificity, high in specificity and the like, and capable of effectively enriching Sca-1 positive adipose-derived mesenchymal stem cells. Specific surface molecular markers such as adherent growth, three-line differentiation capability and high purity; meanwhile, an ADSC-AM method is innovatively used, and the adipose-derived mesenchymal stem cells obtained by the ADSC-AM method are optimal in performance.

Provided are a method of producing a human pluripotent stem cell-derived Leydig-like cell (Leydig-like cells) capable of secreting testosterone stably and persistently, and a human pluripotent stem cell-derived Leydig-like cell. Also provided is a human pluripotent stem cell-derived Leydig-like cell that enables long-term maintenance of a Leydig-like cell secreting a sufficient amount of testosterone, and is improved in efficiency of differentiation induction. The method includes the steps of: culturing a human pluripotent stem cell under a culture condition with addition of one or a plurality of kinds selected from the group consisting of: a WNT canonical pathway activator; bone morphogenetic protein 4 (BMP4); and vascular endothelial growth factor (VEGF); and forcedly expressing NR5A1, and is achieved by controlling the timing of addition and removal of cyclic adenosine monophosphate (cAMP), suspension culture, adhesion culture, and the like.

Provided are a production method of retinal pigment epithelial (RPE) cells that improves differentiation induction efficiency of pluripotent stem cells into RPE cells, and can provide highly pure RPE cells by a simple and easy operation in a short period, a culture method of RPE cells that can stably grow and culture a cell, a toxicity / efficacy evaluation method using RPE cells useful for transplantation therapy, and a therapeutic drug for a retinal disease. The invention relates to a production method of RPE cells, comprising adhesion culture of human pluripotent stem cells using a culture substrate coated with a laminin-E8 fragment, a culture method of RPE cells, comprising adhesion culture of RPE cells using a culture substrate coated with a laminin-E8 fragment, a toxicity or efficacy evaluation method using RPE cells obtained by producing or culturing by the method, and a therapeutic drug for a retinal disease, containing the RPE cells.

To provide methods for inducing differentiation to ectodermal cells excellent in differentiation-inducing efficiency from pluripotent stem cells to ectodermal cells and excellent in culture operability as well as methods for making such cells.SOLUTION: Methods for inducing differentiation from pluripotent stem cells to ectodermal cells comprise the steps of: (1) coating a cell culture substrate with one or more substances selected from the group consisting of Matrigel, laminin, fibronectin, vitronectin and collagen where the substrate comprises a region having cell adhesiveness and cell-proliferating property and a region having neither of them and then seeding pluripotent stem cell 1; (2) conducting adhesion culture of the seeded pluripotent cells in the presence of an undifferentiation-maintaining medium in feeder-free conditions to form agglutination of pluripotent stem cells 2 adhered to the cell culture substrate; (3) culturing the formed agglutination of pluripotent stem cells in the presence of a culture medium containing a differentiation-inducing agent so that the cells adhered to the cell culture substrate form germ layer 3; and (4) culturing the formed germ layer in the presence of a culture medium containing the differentiation-inducing agent in the adhered state to the cell culture substrate to form ectodermal cell aggregates 4.SELECTED DRAWING: Figure 1

The application discloses a method for stably producing exosomes of regenerative repair and anti-inflammatory molecules of sheep umbilical cord mesenchymal stem cells and application thereof in skin care products, and comprises the following steps: S1, using healthy sheep umbilical cord as raw material, obtaining primary sheep umbilical cord mesenchymal stem cells through aseptic treatment, Wharton's jelly extraction and tissue block adherent culture; S2, introducing an immortalization gene into the primary sheep umbilical cord mesenchymal stem cells through a lentivirus transduction technology, and obtaining an immortalized sheep umbilical cord mesenchymal stem cell line through resistance screening; S3, collecting supernatant after amplification culture of the immortalized sheep umbilical cord mesenchymal stem cell line, and purifying through multi-stage centrifugal impurity removal and PEG precipitation method to obtain stably produced exosomes containing regenerative repair and anti-inflammatory molecules. The exosomes containing regenerative repair and anti-inflammatory molecules provided by the application exhibit significant advantages in yield and quality, and have outstanding application value in the fields of medical beauty anti-aging and skin care products.

The present invention provides a method for producing dopaminergic neuron progenitor cells from pluripotent stem cells, which method comprises the steps of: (i) performing adherent culture of pluripotent stem cells on an extracellular matrix in a medium containing a reagent(s) selected from the group consisting of BMP inhibitor, TGFβ inhibitor, SHH signal-stimulating agent, FGF8, and GSK3β inhibitor; (ii) collecting Corin- and / or Lrtm1-positive cells from the cells obtained in Step (i) using a substance which binds to Corin and / or a substance which binds to Lrtm1; and (iii) performing suspension culture of the cells obtained in Step (ii) in a medium containing a neurotrophic factor.

The invention provides a cell chiral growth regulation and control method, and relates to the technical field of cell chiral culture. The regulation and control method provided by the invention comprises the following steps: carrying out full-exposure pre-shaping on a photo-crosslinking substrate, then inoculating cells to be subjected to chiral culture, carrying out adherent culture, then carrying out patterning exposure and continuous culture, and differentiating to obtain a cell chiral array; the photo-crosslinking substrate is provided with photo-crosslinking gel and a matched photoinitiator. In-situ regulation and control on cell chirality can be realized on the basis of the cell chirality growth direction through a double-exposure method, so that cell chirality arrays with different chirality orientations can be obtained after differentiation, and the structure and function of the heart and basic unit myocardial cells thereof can be known and simulated; the method is of great significance to cardiovascular disease research and drug screening.

The invention relates to the technical field of cell culture, in particular to a multi-union bioreactor for adherent culture of cells, which is provided with a rotating mechanism and a culture mechanism, so that multiple groups of cells can be cultured in the same environment with the same temperature and humidity in the cell culture process, and the culture efficiency is improved; comprising multiple groups of culture mechanisms; the device further comprises a heating mechanism, a rotating mechanism, a driving mechanism and a plurality of fixing mechanisms, the rotating mechanism is installed on the heating mechanism, the multiple culture mechanisms are installed on the rotating mechanism, the driving mechanism is installed on the rotating mechanism, and the fixing mechanisms are installed on the rotating mechanism; the heating mechanism is used for heating, the rotating mechanism is used for rotating, the culture mechanism is used for culturing, the driving mechanism is used for driving the culture mechanism, and the fixing mechanism is used for fixing the culture mechanism.

The invention discloses a culture medium for in-vitro adherent culture of primary tumor cells and application, and relates to the technical field of tumor cell culture. Comprising a basic culture medium, fetal calf serum FBS with the concentration of 5 ng / mL-15 ng / mL, an epidermal growth factor EGF with the concentration of 1 ng / mL-10 ng / mL, a basic fibroblast growth factor bFGF with the concentration of 0.5 ng / mL-5 ng / mL, an insulin-transferrin-selenium ITS supplement with the final volume concentration of 0.1%-1%, L-glutamine with the final volume concentration of 0.5 mM-2 mM, ascorbic acid with the final volume concentration of 0.1%-0.5% and non-essential amino acid with the final volume concentration of 1%-5%. The kit comprises 10 [mu] g / mL-50 [mu] g / mL of heparin and 1 [mu] g / mL-5 [mu] g / mL of an Rho kinase inhibitor. The culture medium overcomes the defects that in the prior art, when primary tumor cells are subjected to adherent culture, apoptosis or dedifferentiation is prone to occurring, in-vivo tumor characteristics are difficult to simulate, the tumor growth speed is low, the proliferation effect is poor, and the adherent rate is low.

The invention belongs to the technical field of tissue engineering and aging resistance, and particularly relates to a composition for delaying cell aging and maintaining tissue homeostasis as well as a preparation method and application of the composition. Specifically, research finds that adipose tissues, skin tissues and the like are placed in a serum-free basal culture medium, adherent culture is performed, the obtained supernate is collected, and the obtained serum-free conditioned culture medium preparation can effectively inhibit aging of cells (such as ADSCs or HDFs) and maintain the functional steady state of the cells in in-vitro culture. Meanwhile, the culture medium product provided by the invention is also proved to be capable of inhibiting an IL-6 / STAT3 signal channel, and the channel is revealed to be a core mechanism of the anti-aging effect of the culture medium product, so that mechanism support is provided for the application of the culture medium product to cell aging resistance, and the culture medium product has good practical application value.

Provided are a method of producing a human pluripotent stem cell-derived Leydig-like cell (Leydig-like cells) capable of secreting testosterone stably and persistently, and a human pluripotent stem cell-derived Leydig-like cell. Also provided is a human pluripotent stem cell-derived Leydig-like cell that enables long-term maintenance of a Leydig-like cell secreting a sufficient amount of testosterone, and is improved in efficiency of differentiation induction. The method includes the steps of: culturing a human pluripotent stem cell under a culture condition with addition of one or a plurality of kinds selected from the group consisting of: a WNT canonical pathway activator; bone morphogenetic protein 4 (BMP4); and vascular endothelial growth factor (VEGF); and forcedly expressing NR5A1, and is achieved by controlling the timing of addition and removal of cyclic adenosine monophosphate (cAMP), suspension culture, adhesion culture, and the like.

The present invention relates to a composition for improving the efficiency of cell suspension culture and a composition for improving the efficiency of cell adhesion culture. According to the present invention, it is possible to artificially alter the anchorage dependency of target cells to be cultured and to simply revert the altered phenotype, if necessary. According to the present invention, it is possible to alter the cell phenotype to an optimal state suitable for the culture purpose and environment and revert the altered phenotype to the original phenotype at a desired time point, thereby maximizing the efficiency of culture of various target cells, including host cells for recombinant protein production, as well as therapeutic immune cells and stem cells.

The invention relates to the field of aerospace biology and cell biology, in particular to an in-vitro preparation method of myocardial cells under a supergravity condition. Comprising the following steps: step 1, performing adherent culture on myocardial cells on a two-dimensional matrix; 2, mixing the myocardial cells subjected to two-dimensional culture in the step 1 with matrigel to form a three-dimensional culture system; step 3, placing the three-dimensional culture system obtained in the step 2 in a supergravity environment for culture; wherein the gravity intensity of the supergravity environment is 3G, and the treatment time is 12-96 hours. The invention provides a complete and repeatable myocardial cell in-vitro preparation method under the supergravity condition for the first time. A stable 3G supergravity environment can be provided, the damage of high shear force to cells is effectively reduced, the core pain point that a traditional supergravity experiment depends on expensive and rare parabola flight or a large centrifugal machine is solved, and the research cost and the technical threshold are greatly reduced.

The invention is applicable to the technical field of biological medicine, and provides a metformin-engineered umbilical cord mesenchymal stem cell and a preparation method and application thereof, and the preparation method comprises the following steps: taking fresh umbilical cord tissue, cleaning, cutting into small blocks, performing adherent culture, and performing fusion passage when the cell grows to 80%; and co-incubating the P5-generation human umbilical cord mesenchymal stem cells and metformin, and centrifugally collecting the cells after pancreatin digestion to obtain the metformin-engineered umbilical cord mesenchymal stem cells. The concentration of the metformin is 500 [mu] mol / L. Metformin is used for pretreating human umbilical cord mesenchymal stem cells to construct Met-MSC with metabolism enhancement and function optimization characteristics, so that the Met-MSC has stronger anti-apoptosis, anti-oxidation, angiogenesis promotion and immunoregulation capabilities, a safe, simple, convenient and convertible new cell treatment strategy is provided, and the application bottleneck of traditional MSC treatment in diabetic foot ulcer is broken through.

The invention discloses a method for evaluating in-vitro lymphopoiesis inhibition of mesenchymal stem cells, and relates to the technical field of biological medicine, the method for evaluating in-vitro lymphopoiesis inhibition of mesenchymal stem cells comprises the following steps: S1, preparing an MSC cell suspension; s2, preparing a PBMC (peripheral blood mononuclear cell) suspension, namely preparing a commercialized mixed source PBMC with a built line; s3, MSC inoculation adherent culture is carried out; s4, carrying out CFSE dyeing and inoculation; s5, marking the PBMC proliferative cells by using the MSC: after culture is completed, adding a cell activity detection reagent, incubating in a dark place, detecting, and carrying out statistical analysis; according to the method, the sample size of the same batch of PBMCs is increased by adopting commercialized established-line mixed source PBMCs, so that the difference between batches is reduced; compared with a traditional evaluation method, the method provided by the invention can more comprehensively and reliably evaluate the in-vitro lymphocyte proliferation inhibition capability, so that a basis is provided for clinical application of mesenchymal stem cells.

The utility model discloses a bioreactor capable of realizing large-scale adherent culture of microalgae, which belongs to the technical field of microalgae culture equipment and comprises a support system, an adherent culture unit and a culture medium circulating module. The culture medium circulating module comprises a circulating water tank, a circulating pipeline and a liquid supplementing branch pipe which are communicated in sequence; the support system comprises a main support and support branches connected to the two opposite sides of the main support at equal intervals, each support branch is covered with a supporting body, the culture carrier is hung on the supporting bodies, and a fixing clamp used for enabling the two half parts of the culture carrier to be attached together is arranged on the culture carrier. According to the utility model, the adherent culture units are convenient to disassemble and assemble, the liquid supplementing branch pipes are convenient to protect, and part of the liquid supplementing branch pipes can be selectively selected for infusion work, so that the culture scale is convenient to adjust; and more adherent culture units can be arranged in the same space.

The application discloses a culture dish and its use in preparing a cell sheet. The culture dish comprises a body defining a culture space inside, and a coating arranged at the bottom of the culture space, wherein the surface free energy of the coating is not more than 90 mJ / m 2 . Thus, by adjusting the ion concentration in the cell liquid medium, the binding force between the coating and the cells can be reduced, so that the adherent cultured cells can be detached from the coating at the bottom of the culture dish in the form of a cell sheet, and the cell sheet is effectively obtained.