CN109694825A - A kind of mold for cultivating 3D cell ring - Google Patents

A kind of mold for cultivating 3D cell ring Download PDF

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Publication number
CN109694825A
CN109694825A CN201910126589.2A CN201910126589A CN109694825A CN 109694825 A CN109694825 A CN 109694825A CN 201910126589 A CN201910126589 A CN 201910126589A CN 109694825 A CN109694825 A CN 109694825A
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China
Prior art keywords
tubular column
open tubular
die holder
cell
cell ring
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CN201910126589.2A
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Chinese (zh)
Inventor
吴宏伟
黄钢
钟午
许彦
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Hunan Cancer Hospital
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Hunan Cancer Hospital
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Priority to CN201910126589.2A priority Critical patent/CN109694825A/en
Publication of CN109694825A publication Critical patent/CN109694825A/en
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/02Form or structure of the vessel
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/02Form or structure of the vessel
    • C12M23/12Well or multiwell plates
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/48Holding appliances; Racks; Supports

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  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Clinical Laboratory Science (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The invention discloses a kind of molds for cultivating 3D cell ring, including lower die holder and upper die holder, the lower die holder include chassis, and the chassis is disc, are provided centrally with the rotary table to raise upward, are provided with central rods above the rotary table;The upper die holder includes open tubular column and bracket, and the bracket shares two sections, is fixed on the periphery of the open tubular column, and two sections of brackets are symmetrical centered on the open tubular column, the either flush of the rack upper surface and the open tubular column.Beneficial effect is: for manufactured cell ring naked eyes as it can be seen that internal diameter reaches 2mm, extracellular matrix is abundant, can clamp cell ring, manually convenient for observation and subsequent experimental operation;Mold design is succinct, easy-to-use, and strong operability is at low cost after large-scale production, and with existing culture board size perfect matching, scalability is strong, high-volume can complete the preparation of cell ring simultaneously.

Description

A kind of mold for cultivating 3D cell ring
Technical field
The present invention relates to biological cell culture fields, and in particular to a kind of mold for cultivating 3D cell ring.
Background technique
In vitro study in terms of conventional cell biological study and disease treatment mostly uses two-dimentional cell culture mode, institute The culture plane of offer is flat.However under physiological status, histocyte is to grow in three dimensions, cell and cell Between not only contact with each other, but also to be in contact with support constructions such as extracellular matrixs.Two-dimentional cell culture mode can not be true The positive extracellular matrix embodied in tissue, therefore many physiological reactions in this mode, such as expression of receptor, transcriptional expression, cell What happens migrating and apoptosis etc., being proved from practical organ or tissue is different.Moreover, cell from division, be proliferated, move It moves to apoptosis, whole process depends on its internal intrinsic solid space and temporal organizational principle by accuracy controlling.Two dimension Also there is a big difference for function of the cell culture mode from accurate reproduction tissue inner cell.
In vivo, cell is grown in the environment of a 3D, i.e., by other cells, membranous system, fibrous layer, extracellular matrix Equal tight encirclements.Studies have shown that the cell grown in 3D culture systems can be shown closer to physiological phenotype, interaction And reaction, it can more accurately reappear cell behavior.Many researchs are concerned about, and the conclusion obtained using dimensional culture and animal are real Test similar, Three-dimensional cell culture can more accurately simulate normal cellular morphology, proliferation, differentiation and migration, therefore study Three-dimensional cell culture technology is particularly important.Currently, in vitro in biomedical research, from 2D to the cell culture system of 3D Transformation gradually carries out.In addition, the detection carried out under external 3D cell culture system can be used as internal animal model and verify it A kind of preceding alternative greatlys save cost and the period of experiment in vivo.
Applicants have discovered that at least there is following technical problem in the prior art: dimensional culture technology, which is broadly divided into, at present needs It wants the dimensional culture technology of bracket and does not need the Three-dimensional cell culture technology of bracket.Cytoskeleton is mainly in three-dimensional space The porous structure for adhering to for cell and growing is constructed, cell depends on bracket and carries out three dimensional growth and migration, main bracket material Material has collagen and hydrogel etc.;The dimensional culture technology for not needing bracket, which mainly passes through physical method, is suspended in attached cell To achieve the purpose that dimensional culture in culture medium, current main technology has microcarrier, magnetic suspension, hanging-drop plates and magnetic three-dimensional raw The technologies such as object printing, complicated operation for such methods, and early investment is larger, and overwhelming majority dimensional culture is by means of branch at present Frame material.But the space occupy-place of gel or bracket influences the three-dimensional structure of cell formation rule densification, catabolite is also right The Growth and Differentiation of cell has an impact, and higher cost is also its unfavorable factor, is not still ideal dimensional culture strategy.They Mostly there is also operating process complexity, bracket influences cell Growth and Differentiation, and culture cell volume is less than normal, and subsequent operation inconvenience etc. is asked Topic, from ideal 3D cell culture model, there is also biggish gaps, and current product produces greatly mostly from external company Product are expensive, limit its extensive use at home, hinder the development of domestic medical research.
Summary of the invention
The object of the invention is that a kind of mold for cultivating 3D cell ring is provided to solve the above-mentioned problems, to solve 3D cell culture is at high cost in the prior art, the technical problems such as complicated for operation.In many technical solutions provided by the invention preferably Technical solution include the technical effects such as structure is simple, at low cost and easier to operate, elaboration as detailed below.
To achieve the above object, the present invention provides following technical schemes:
The present invention provides a kind of molds for cultivating 3D cell ring, including lower die holder and upper die holder, the lower die holder to include Chassis, the chassis are disc, are provided centrally with the rotary table to raise upward, are provided with central rods above the rotary table;
The upper die holder includes open tubular column and bracket, and the bracket shares two sections, is fixed at the open tubular column On periphery, and two sections of brackets are symmetrical centered on the open tubular column, the rack upper surface and the open tubular column Either flush;
The upper die holder is slidably socketed on the rotary table, and the central rods protrude into the open tubular column, to make described An annular growing space is formed between open tubular column and the central rods, the annular growing space is for top opening and with described Rotary table upper surface is the circular column space of planes of bottom surface.
Using a kind of above-mentioned mold for cultivating 3D cell ring, the chassis outer diameter of the lower die holder and the branch of the upper die holder Length between frame both ends is consistent with the culture hole aperture on porous cell culture plate, and wherein porous cell culture plate can be 6 holes, 12 holes, 24 holes, 48 holes, 96 holes etc., the chassis is put into the culture hole of porous cell culture plate, is used to support and is stablized The open tubular column of the upper die holder, the central rods play compartmentation, a top opening are combined into together with the open tubular column, high The annular growing space of 4mm, cell is self-service in the annular growing space to dress up ring-type, the institute at the top of the open tubular column Bracket is stated to be in contact with the culture hole inner wall of porous cell culture plate, thus be to be stable at culture hole center by the open tubular column, After initial stage 3D cell ring culture molding, it can remove upper die holder, cell ring made to continue to cultivate in lower die holder, it is thin after taking-up upper die holder More fully nutrition and oxygen in the available culture solution of born of the same parents' ring, convenient for being trained stable cell ring and taking out the subsequent place of progress Reason;The upper die holder of upgrading, which can also improve, becomes the cylindrical wall with semi-transparent function, experimental can need in cylindrical wall Outer culture solution adds different inducible factors and carries out observation processing to cell ring;Cultivate molding cell ring can be taken off and other Cell ring carries out tandem compound, to be superimposed as tubular fossils pipe.
Preferably, the central rods are the hemispherical cylindrical body in top.
Preferably, the radius on the chassis is 7.7mm, high 1mm, the radius of the rotary table is 2mm, high 1.5mm, described The radius of central rods is 1mm, and is 7mm to the spacing between the chassis bottom surface at the top of the central rods.
Preferably, the upper surface of the rotary table is the high outside and low outside curved surface with radian, and the radian of the curved surface is 0.125°。
Preferably, the outer diameter of the open tubular column is 3mm, internal diameter 2mm, high 6mm, the unilateral stent length is 4.7mm, high 2mm, wide 1mm.
Preferably, the outer edge of the bracket is and the circular arc using the center of the open tubular column as the arc surface in the center of circle The radius in face is 7.7mm.
Preferably, the volume of the annular growing space is 37.68 μ l.
The utility model has the advantages that cell ring naked eyes made of the 1, present invention are as it can be seen that internal diameter reaches 2mm, extracellular matrix is abundant, can be empty Hand clamps cell ring, convenient for observation and subsequent experimental operation;
2, mold design is succinct, easy-to-use, and strong operability is at low cost after large-scale production, perfect with existing culture board size Matching, scalability is strong, high-volume can complete the preparation of cell ring simultaneously.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this Some embodiments of invention for those of ordinary skill in the art without creative efforts, can be with It obtains other drawings based on these drawings.
Fig. 1 is space diagram of the invention;
Fig. 2 is cross-section diagram of the invention;
Fig. 3 is lower die holder space diagram of the invention;
Fig. 4 is upper die holder space diagram of the invention.
The reference numerals are as follows:
1, lower die holder;101, chassis;102, rotary table;103, central rods;2, upper die holder;201, open tubular column;202, bracket;3, Annular growing space.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, technical solution of the present invention will be carried out below Detailed description.Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Base Embodiment in the present invention, those of ordinary skill in the art are obtained all without making creative work Other embodiment belongs to the range that the present invention is protected.
Referring to shown in Fig. 1-4, the present invention provides a kind of molds for cultivating 3D cell ring, including lower die holder 1 and upper die holder 2, the lower die holder 1 includes chassis 101, and the chassis 101 is disc, convenient for enabling lower die holder 1 to be put into existing cell training In the culture hole for supporting plate, chassis 101 is provided centrally with the rotary table 102 to raise upward, is provided with central rods above the rotary table 102 103;The upper die holder 2 includes open tubular column 201 and bracket 202, and the bracket 202 shares two sections, is fixed at the sky On the periphery of stem 201, and two sections of brackets 202 are symmetrical centered on the open tubular column 201, convenient for making upper die holder 2 are socketed on the rotary table 102 of lower die holder 1 by open tubular column 201, and are positioned by bracket 202 to open tubular column 201, thus So that open tubular column 201 is kept coaxial with central rods 103, at the same make mold in the culture hole for being put into tissue culture plate when, pass through branch Frame 202 is supported at the top of cell culture well, to improve the stability of mold placement, 202 upper surface of bracket and the sky The either flush of stem 201 simplifies its processing technology convenient for the processing of upper die holder 2, reduces processing cost;The upper die holder 2 is sliding Dynamic to be socketed on the rotary table 102, the central rods 103 protrude into the open tubular column 201, thus make the open tubular column 201 with An annular growing space 3 is formed between the central rods 103, the annular growing space 3 is for top opening and with the circle 102 upper surface of platform is the circular column space of planes of bottom surface, and compared with the gel of the prior art and bracket, it is empty to be both not take up cell growth Between, so that cell is capable of forming the three-dimensional structure of dense regular, while any catabolite will not be generated, therefore will not influence thin Intracellular growth differentiation facilitates later observations and experimental implementation so that the cell volume of culture be made to be capable of increasing.
As this case preferred embodiment, the central rods 103 are the hemispherical cylindrical body in top, so set, The cell ring for being capable of forming circular ring shape in culture convenient for cell facilitates observation and experiment so as to manually clamp cell ring; The radius on the chassis 101 is 7.7mm, high 1mm, and the radius of the rotary table 102 is 2mm, high 1.5mm, the central rods 103 Radius is 1mm, and is 7mm to the spacing between 101 bottom surface of chassis at the top of the central rods 103, so set, being convenient for So that lower die holder 1 is put into the culture hole of culture plate completely, while guaranteeing that chassis 101 fits with culture hole inner wall, makes thin In born of the same parents' incubation, avoid the level in tissue culture plate of lower die holder 1 that from sliding, upper die holder 2 is injection moulded using polystyrene, Polystyrene is a kind of colorless and transparent thermoplastic plastic, can so keep the lens degree of upper die holder 2, convenient for thin inside observation Intracellular growth state, while can be for further processing to the surface of upper die holder 2, to increase the hydrophobicity on 2 surface of upper die holder, to have Conducive to cell adhesion on the side wall of central rods 103, and in cell cultivation process, cell growth is done in no magnetic field It disturbs, so as to turn out the cell ring for having more stereoscopic features more greatly than the microsphere volume of Hanging drop culture, consequently facilitating operation, institute The upper surface for stating rotary table 102 is the high outside and low outside curved surface with radian, and the radian of the curved surface is 0.125 °, so set, just In pooling together the cell of annular 3 bottom of growing space, while cell is given with the bottom space of self assembly cyclization.
The outer diameter of the open tubular column 201 is 3mm, internal diameter 2mm, high 6mm, and unilateral 202 length of the bracket is 4.7mm, high 2mm, wide 1mm, so set, convenient for enabling open tubular column 201 to be socketed on rotary table 102, while making open tubular column 201 Top it is concordant with the top of central rods 103, to central rods 103 are all placed in the inner hole of open tubular column 201, so that annular The height of growing space 3 reaches maximum, to improve the culture height of 3D cell ring, polystyrene note is can be used in open tubular column 201 Molding is penetrated, had so both guaranteed the transparency of open tubular column 201, from being carried out from outside to internal cell, while convenient for logical It crosses open tubular column 201 and keeps apart annular growing space 3 with culture hole, thus guarantee that annular growing space 3 has Packed surface, Open tubular column 201 can also be used polycarbonate film and the semi-transparent cylindrical body with semi-permeable membrane function be made, and can so facilitate open tubular column The cell culture fluid in 201 outsides enters central annular growing space 3 by semi-permeable membrane, or convenient for the cell inside and outside semi-permeable membrane because The exchange of son or cell active substance, consequently facilitating control substance passes in and out annular growing space 3, neither allow utility arbitrarily Exudation, does not also allow harmful substance to enter easily.
The outer edge of the bracket 202 is and the arc surface using the center of the open tubular column 201 as the arc surface in the center of circle Radius is 7.7mm, so set, convenient for making 202 outside of bracket fit with culture hole inner wall, to pass through 202 pairs of skies of bracket The top of stem 201 is supported positioning, i.e., guarantee open tubular column 201 and central rods 103 are coaxial, and open tubular column 201 is avoided to train It supports in hole and shakes, improve the stability that upper die holder 1 is installed;The volume of the annular growing space 3 is 37.68 μ l, compared to existing Technology is suspended in attached cell in culture medium to reach dimensional culture purpose by physical method, and the annular growth of big volume is empty Between can guarantee the diameter and height of cultivated cell ring, so as to which the cell ring that macroscopic internal diameter reaches 2mm, side is made Just manually clamping.
Using the above structure, 202 both ends of bracket of 101 outer diameter of chassis of the lower die holder 1 and the upper die holder 2 it Between length it is consistent with the culture hole aperture on porous cell culture plate, wherein porous cell culture plate can for 6 holes, 12 holes, 24 holes, 48 holes, 96 holes etc., the chassis 101 are put into the culture hole of porous cell culture plate, are used to support and stablize on described The open tubular column 201 of die holder 2, the central rods 103 play compartmentation, are combined into a top together with the open tubular column 201 and open Mouthful, the annular growing space 3 of high 4mm, cell is self-service in the annular growing space 3 to dress up ring-type, the open tubular column The bracket 202 at 201 tops is in contact with the culture hole inner wall of porous cell culture plate, to be by the open tubular column 201 Be stable at culture hole center can remove upper die holder 2, continues cell ring in lower die holder 1 after initial stage 3D cell ring culture molding More fully nutrition and oxygen in the available culture solution of cell ring after upper die holder 2 is taken out in culture, stable thin convenient for being trained Born of the same parents' ring and taking-up carry out subsequent processing;The upper die holder 2 of upgrading, which can also improve, becomes the cylindrical wall with semi-transparent function, can basis Experiment needs to add different inducible factors to the culture solution inside and outside cylindrical wall and carries out observation processing to cell ring;Culture molding Cell ring can be taken off with other cell rings carry out tandem compound, to be superimposed as tubular fossils pipe.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain Lid is within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.

Claims (7)

1. a kind of mold for cultivating 3D cell ring, it is characterised in that: including lower die holder (1) and upper die holder (2), the lower die holder It (1) include chassis (101), the chassis (101) is disc, is provided centrally with the rotary table (102) to raise upward, the rotary table (102) top is provided with central rods (103);
The upper die holder (2) includes open tubular column (201) and bracket (202), and the bracket (202) shares two sections, is fixedly installed On the periphery of the open tubular column (201), and two sections of brackets (202) are symmetrically divided centered on the open tubular column (201) Cloth, the either flush of bracket (202) upper surface and the open tubular column (201);
The upper die holder (2) is slidably socketed on the rotary table (102), and the central rods (103) protrude into the open tubular column (201) It is interior, to make to form an annular growing space (3), the annular between the open tubular column (201) and the central rods (103) Growing space (3) is top opening and using the rotary table (102) upper surface as the circular column space of planes of bottom surface.
2. a kind of mold for cultivating 3D cell ring according to claim 1, it is characterised in that: the central rods (103) are top The hemispherical cylindrical body in portion.
3. a kind of mold for cultivating 3D cell ring according to claim 2, it is characterised in that: the radius of the chassis (101) For 7.7mm, high 1mm, the radius of the rotary table (102) is 2mm, high 1.5mm, and the radius of the central rods (103) is 1mm, and The spacing arrived between the chassis (101) bottom surface at the top of the central rods (103) is 7mm.
4. a kind of mold for cultivating 3D cell ring according to claim 3, it is characterised in that: the upper table of the rotary table (102) Face is the high outside and low outside curved surface with radian, and the radian of the curved surface is 0.125 °.
5. a kind of mold for cultivating 3D cell ring according to claim 4, it is characterised in that: outside the open tubular column (201) Diameter is 3mm, internal diameter 2mm, high 6mm, and the unilateral bracket (202) length is 4.7mm, high 2mm, wide 1mm.
6. a kind of mold for cultivating 3D cell ring according to claim 5, it is characterised in that: the outside of the bracket (202) Edge be using the center of the open tubular column (201) as the arc surface in the center of circle, and the radius of the arc surface be 7.7mm.
7. a kind of mold for cultivating 3D cell ring according to claim 6, it is characterised in that: the annular growing space (3) Volume be 37.68 μ l.
CN201910126589.2A 2019-02-20 2019-02-20 A kind of mold for cultivating 3D cell ring Pending CN109694825A (en)

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CN113528343A (en) * 2021-07-19 2021-10-22 中国科学院重庆绿色智能技术研究院 Adherent cell culture device for terahertz wave irradiation
CN113528343B (en) * 2021-07-19 2022-08-02 中国科学院重庆绿色智能技术研究院 Adherent cell culture device for terahertz wave irradiation

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