Stem cell in-vitro culture device based on hollow columnar three-dimensional scaffold material

Abstract

The utility model discloses a stem cell in-vitro culture device based on a hollow columnar three-dimensional support material, which comprises a culture box, the front side of the culture box is of an opening structure, a plurality of rotating seats are longitudinally distributed in the culture box, a mounting rack is fixedly connected among the rotating seats, a culture dish is arranged in the mounting rack, and the culture dish is connected with the culture box. A hollow columnar three-dimensional bracket is arranged in the culture dish, a porous structure is arranged in the hollow columnar three-dimensional bracket, and stem cells are arranged in the hollow columnar three-dimensional bracket. Compared with the prior art, the hollow columnar three-dimensional scaffold material has the advantages that exosomes can be easily collected in the stem cell culture process, the hollow columnar three-dimensional scaffold material can achieve large-scale amplification on the number of cells, the hollow columnar three-dimensional scaffold material can be degraded, and the stem cells can be harvested in a lossless mode.

Description

TECHNICAL FIELD

[0001] The utility model relates to stem cell culture technical field, concretely is a stem cell in vitro culture device based on hollow columnar three -dimensional support material. BACKGROUND

[0002] The high-quality, standardized and scaled in-vitro expansion culture of stem cells is always a core technical difficulty to be overcome. At present, the common way of in-vitro culture of most stem cells is in traditional two-dimensional cell culture plate / dish, relying on specific culture medium, cytokines and small molecules to support the growth of stem cells. However, this traditional two-dimensional monolayer or multilayer plate culture, similar to the multilayer culture in the laboratory, is mainly a "cell factory" with high consumption and low output, which consumes a lot of manpower and material resources, and it is difficult to maintain the stability between batches of stem cells. Stem cell related technology products require cells to have consistency (i.e. low batch difference), high quality, mass production and low cost, etc. However, stem cells are extremely sensitive to the surrounding microenvironment during in-vitro culture, which includes 3D spatial structure, biochemical factors, pH value, dissolved oxygen, fluid shear force of culture solution, and concentration of metabolites in culture medium. These microenvironment compositions play a crucial role in stem cell expansion culture, but existing domestic and foreign related technology products have not yet achieved large-scale expansion culture of cells (stem cells) through cell microenvironment design.

[0003] In order to construct a similar three-dimensional internal environment of stem cells in vitro as in vivo, research on in-vitro three-dimensional culture has gradually increased, and different types of three-dimensional systems have been gradually developed and utilized. The most common are hydrogel three-dimensional systems and scaffold three-dimensional culture systems, such as hydrogel systems constructed based on polyethylene glycol, sodium alginate, hyaluronic acid, mouse myeloma extracellular matrix components, and scaffold systems constructed using gelatin, collagen, polyethylene glycol, sodium alginate, hyaluronic acid, etc. Both of them can achieve in-vitro three-dimensional culture of cells.

[0004] Chinese patent application No. 201110375284.9 discloses a method and device for constructing a three-dimensional microenvironment, which dissolves polymer monomer polyethylene glycol diacrylate and 2-hydroxy-4-(2-hydroxyethoxy)-2-methyl propiophenone in a saturated NaCl solution to obtain a photo-crosslinkable prepolymer solution, adds NaCl particles with a diameter of 100-125 μm as a pore-forming agent to the photo-crosslinkable prepolymer solution, and irradiates and crosslinks the solution by using a UV light source to obtain a scaffold material. Although the scaffold material has good biocompatibility and mechanical properties, it is non-degradable, so that cells cannot be collected after the cells are cultured, and the cells cannot be subcultured. Even if the cells can be harvested, the number of cells is very small and cannot meet the needs of downstream biochemical detection and future clinical needs of cell therapy. Another problem is that the degree of photo-crosslinking changes with the energy of the light source and the irradiation distance, so it is difficult to maintain the same degree of crosslinking in different batches of scaffolds.

[0005] Through retrieval, it is found that a patent (Patent No. ZL201280042758.0) granted in 2016 uses chemical crosslinking and photo-crosslinking methods respectively to construct a three-dimensional system with polyethylene glycol diacrylate as the main body. Whether chemical crosslinking or photo-crosslinking, the crosslinking process is irreversible, and the three-dimensional system constructed is non-degradable, so that cells in the three-dimensional scaffold cannot be all harvested without damage. Such a culture system is also not conducive to the adhesion of cells and the scaffold, resulting in difficulty in adherent growth of stem cells.

[0006] The information disclosed in this BACKGROUND section is only for the purpose of increasing the understanding of the general background of the application, and should not be taken as an acknowledgment or any form of suggestion that this information forms prior art that is known to those of ordinary skill in the art. Practical new type content

[0007] The technical problem to be solved by the present application is to overcome the above technical defects, and to provide a stem cell in vitro culture device based on a hollow columnar three-dimensional scaffold material.

[0008] In order to solve the above problems, the technical scheme of the present application is as follows: a stem cell in vitro culture device based on a hollow columnar three-dimensional scaffold material, comprising a culture box, the front side of the culture box is an open structure, a plurality of rotating seats are longitudinally distributed in the culture box, a mounting rack is fixedly connected between the plurality of rotating seats, a culture dish is arranged in the mounting rack, and a hollow columnar three-dimensional scaffold is arranged in the culture dish. The interior of the hollow columnar three-dimensional scaffold is provided with a porous structure, and the interior of the hollow columnar three-dimensional scaffold is provided with stem cells.

[0009] Further, the hollow columnar three-dimensional scaffold is easy to collect exosomes during the culture of stem cells.

[0010] Further, the hollow cylindrical three-dimensional scaffold material can be degraded, and the stem cells are harvested.

[0011] Further, the hollow cylindrical three-dimensional scaffold material is a sponge-like three-dimensional biomaterial.

[0012] The utility model has the advantages compared with prior art:

[0013] 1, hollow cylindrical three-dimensional scaffold material of stem cell in vitro culture, increase cell inoculation and culture area, and the inner surface and the outer surface can satisfy cell growth, improve culture effect, and save culture medium.

[0014] 2, hollow cylindrical three-dimensional scaffold material of stem cell in vitro culture inoculates cell simply, need not single inoculation, and the material is a sponge-like three-dimensional biomaterial, mainly relies on the active siphon action of material, and cell can be inoculated in three-dimensional material, and cell distribution is more uniform.

[0015] 3, three-dimensional scaffold material is internally porous structure, and is closer to real physiological environment in vivo.

[0016] 4, hollow cylindrical three-dimensional scaffold material is easy to collect exosome in the process of culturing stem cell, and the number of cell can be realized scale expansion in hollow cylindrical material, and hollow cylindrical three-dimensional scaffold material can be degraded, and stem cell is harvested without damage. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 It is the three-dimensional structure of the incubator of the utility model Figure One .

[0018] Figure 2 It is the three-dimensional structure of the incubator of the utility model Figure Two .

[0019] Figure 3 It is the hollow cylindrical three-dimensional scaffold material of the utility model.

[0020] Figure 4 It is the cultivation of hollow cylindrical three-dimensional scaffold material inoculated cell in culture medium of the utility model.

[0021] Figure 5 It is the fluorescent imaging representation result of stem cell growth in hollow cylindrical three-dimensional scaffold material along with time of the utility model.

[0022] Figure 6 It is the fluorescent imaging representation result of stem cell growth on the inner surface of hollow cylindrical three-dimensional scaffold material along with time of the utility model.

[0023] As shown in the figure: 1, incubator;2, rotating seat;3, mounting bracket;4, culture dish;5, hollow cylindrical three-dimensional scaffold;6, sliding groove. DETAILED DESCRIPTION

[0024] The specific embodiments of the present application will be further described below with reference to the drawings. The same parts are denoted by the same reference numerals.

[0025] It should be noted that the words "front", "back", "left", "right", "upper" and "lower" used in the following description refer to the directions in the drawings, and the words "inner" and "outer" refer to the directions towards or away from the geometric center of a particular component.

[0026] In order to make the content of the present application more easily understood, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application.

[0027] As shown in Figures 1 to 6 A stem cell in-vitro culture device based on hollow columnar three-dimensional scaffold material, comprising a culture box 1, the front side of the culture box 1 is an open structure, which creates a culture environment for stem cells through the culture box 1, a plurality of rotating seats 2 are longitudinally distributed in the culture box 1, a mounting rack 3 is fixedly connected between the plurality of rotating seats 2, a culture dish 4 is arranged in the mounting rack 3, a sliding groove 6 is arranged between the mounting racks 3, the culture dish 4 slides in the movable groove 6, and the movable groove 6 and the culture dish 4 are used in cooperation to facilitate the taking of stem cells cultured in the culture dish, a hollow columnar three-dimensional scaffold 5 is arranged in the culture dish 4, a porous structure is arranged in the hollow columnar three-dimensional scaffold 5, the porous structure is more close to the real physiological environment in the body, stem cells are arranged in the hollow columnar three-dimensional scaffold, the number of stem cells is very small, and the separation and purification are difficult, therefore, the stem cells cultured through the platform should be able to expand a certain scale number, and then meet the downstream application, stem cell derivatives exosomes are also proved to have important regulation function on the body, and the platform which can repeatedly and highly produce exosomes and is easy to harvest is also important for the whole stem cell production process, therefore, the hollow columnar three-dimensional scaffold material is easy to collect exosomes in the process of culturing stem cells, the hollow columnar material can realize large-scale expansion in the number of cells, the hollow columnar three-dimensional scaffold material can be degraded, and the stem cells can be harvested without damage.

[0028] The hollow columnar three-dimensional scaffold is easy to collect exosomes in the process of culturing stem cells, the hollow columnar three-dimensional scaffold material for culturing stem cells in vitro, increases the cell inoculation and culture area, the inner surface and the outer surface can both satisfy cell growth, improves the culture effect, saves the culture medium, the hollow columnar three-dimensional scaffold material can be degraded, and stem cells are harvested, the hollow columnar three-dimensional scaffold material for culturing stem cells in vitro inoculates cells simply, and single inoculation is not needed, the material is a three-dimensional biological material with a sponge-like fluffy structure, cells can be inoculated in the three-dimensional material mainly by the active siphon effect of the material, the cells are uniformly distributed, the surface area of stem cell inoculation and culture in vitro can be increased, the cells are more fully contacted with the culture medium, and the culture medium is used to the maximum extent, and the culture medium cost is saved,

[0029] It should be noted that the relational terms herein such as first and second and the like are used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any such actual relationship or order between such entities or actions. Moreover, the terms "comprises", "comprising", or any other variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but can include other elements not expressly listed or inherent to such process, method, article, or apparatus.

[0030] Although the embodiments of the present application have been shown and described, it is to be understood that for the purpose of the present application, the embodiments change, modify, replace, and vary in many ways without departing from the principles and spirit of the present application, and the scope of the present application is defined by the appended claims and their equivalents.

[0031] The above describes the present application and its embodiments, which is not restrictive, and the embodiments shown in the drawings are only one of the embodiments of the present application, and the actual structure is not limited thereto. In general, if a person skilled in the art is inspired by the above description, without departing from the creative purpose of the present application, similar structure and embodiments can be designed without creative design, which should belong to the protection scope of the present application.

Claims

1. A stem cell culture device in vitro based on a hollow cylindrical three-dimensional scaffold material, characterized by: Include incubator (1), the front side of incubator (1) is open structure, several rotating seats (2) are distributed longitudinally in incubator (1), several rotating seats (2) are fixedly connected with mounting rack (3), culture dish (4) is arranged in mounting rack (3), hollow cylindrical three-dimensional support (5) is arranged in culture dish (4), the inside of hollow cylindrical three-dimensional support (5) is provided with porous structure, stem cells are arranged in the inside of hollow cylindrical three-dimensional support.

2. The device for culturing stem cells in vitro based on hollow cylindrical three-dimensional scaffold material according to claim 1, characterized in that: Sliding groove (6) is formed between mounting rack (3), and culture dish (4) is slid in sliding groove (6).

3. The device for culturing stem cells in vitro based on hollow cylindrical three-dimensional scaffold material according to claim 1, characterized in that: The hollow cylindrical three-dimensional support (5) is easy to collect exosomes during the cultivation of stem cells.

4. The device for culturing stem cells in vitro based on hollow cylindrical three-dimensional scaffold material according to claim 1, characterized in that: The material of the hollow cylindrical three-dimensional support (5) can be degraded, and stem cells can be harvested.

5. The device for culturing stem cells in vitro based on hollow cylindrical three-dimensional scaffold material according to claim 1, characterized in that: The hollow cylindrical three-dimensional support (5) is a sponge-like fluffy three-dimensional biomaterial.

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