CN218089628U - Cell culture 3D prints support - Google Patents
Cell culture 3D prints support Download PDFInfo
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- CN218089628U CN218089628U CN202222024383.9U CN202222024383U CN218089628U CN 218089628 U CN218089628 U CN 218089628U CN 202222024383 U CN202222024383 U CN 202222024383U CN 218089628 U CN218089628 U CN 218089628U
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Abstract
The utility model discloses a cell culture 3D prints support, its characterized in that: print integrated into one piece's support body including 3D, be equipped with the multiunit cell culture hole on the support body, the both ends in cell culture hole are linked together with the outer wall of support body respectively. The utility model provides high cell culture efficiency and quality do benefit to the biological activity of arousing the cell, make cell biology characteristic expression more abundant.
Description
Technical Field
The utility model relates to a cell culture field especially relates to a cell culture 3D prints support.
Background
At present, cell culture is carried out in 2D state, such as culture bottles, culture dishes and the like, internationally, and the cells are supported to grow on a two-dimensional plane. In order to increase the activity of cells, culture dishes coated with bioactive materials are provided by high-end foreign brands and all adopt 2D culture, and the 2D culture cannot simulate the microenvironment of a human body, so that the biological characteristics of the cells cannot be fully expressed, and the functionality of the cells is greatly limited. Although many scholars at home and abroad think that the traditional 2D adherent cell culture method is simple and easy to implement, the plane growth environment provided by the adherent culture method is greatly different from the microenvironment for the cells to grow in organisms, and the original form of the cell growth is changed, so that the biological characteristics of the cells cannot be fully expressed.
Disclosure of Invention
The utility model aims at providing a cell culture 3D prints support through using this support, has given the better environment of cell culture, effectively stimulates the biological activity of cell, improves and cultivates efficiency and cultivation effect, makes the cell biology characteristic expression of cultivateing more abundant.
In order to achieve the above purpose, the utility model adopts the technical scheme that: the utility model provides a cell culture 3D prints support, includes the support body, the support body prints integrated into one piece structure for 3D, be equipped with multiunit cell culture hole on the support body, the both ends in cell culture hole respectively with the outer wall of support body is linked together.
In the technical scheme, the outer wall of the support body is provided with the convex block and the groove, and the convex block can be clamped in the groove of the adjacent support body.
In the technical scheme, two groups of convex blocks and two groups of grooves are respectively arranged on the outer wall of the support body, the two groups of convex blocks are respectively arranged on two adjacent side walls of the support body, the two groups of grooves are respectively arranged on the other two adjacent side walls of the support body, and each group of grooves is arranged on the opposite sides of one group of convex blocks.
In the technical scheme, the length of the bracket body is 5-80 mm, the width of the bracket body is 5-80 mm, and the height of the bracket body is 3-30 mm; the aperture of the bacterial culture hole is 100-800 microns.
In the technical scheme, the porosity of the bracket body is 50-90%.
In the above technical scheme, the stent body is provided with a coating, and the coating is arranged on the outer wall of the stent body and the inner wall of the cell culture hole.
In the above technical scheme, the coating is one or more of collagen, methacrylated collagen, poly-D-lysine, silk fibroin, cellular fibronectin, laminin, vitronectin, gelatin, and methacrylated gelatin.
In the above technical solution, the stent body is made of a metal material or a non-metal material, and the metal material is one or more of titanium, titanium alloy, tantalum metal, stainless steel and cobalt-based alloy; the non-metallic material is one or more of polyethylene glycol and polymers thereof, polyethylene glycol diacrylate, collagen, methacrylate collagen, gelatin, methacrylate gelatin, alumina, zirconia, polyether ether ketone, hydroxyapatite, tricalcium phosphate and silicate nano clay.
In order to achieve the above purpose, the utility model discloses a cell culture method of cell culture 3D printing support, its step is:
(1) integrally printing and molding the designed digital support model by using a digital 3D printer to obtain a preformed support body;
(2) cleaning and drying the preformed support body;
(3) sterilizing the preformed support body in the step (2) to obtain a formed support body;
(4) placing the formed bracket body into a culture dish, a culture plate or a culture bottle, and dripping cells and a culture medium to be amplified on the formed bracket body by using a dropper to ensure that the formed bracket body is fully immersed by the culture medium;
(5) putting a culture dish, a culture plate or a culture bottle which is provided with the formed bracket body and the cells into a carbon dioxide incubator for culture and periodically replacing a culture medium;
(6) and (4) washing after the cell culture achieves the use purpose, and collecting the cells.
According to the technical scheme, before the preformed support body is sterilized, the preformed support body is immersed in the solution of the coating material, the preformed support body is kept stand for 4-24 hours, then the preformed support body is taken out, dried and coated with the coating, and after the coating is coated, the sterilization treatment in the step (3) is carried out.
Among the above-mentioned technical scheme, after cell culture reached the use purpose, add specific enzyme earlier to the shaping support body or the shaping support body that has the coating for coating or shaping support body are carried out the enzymolysis by specific enzyme, and coating or shaping support body are by the cell nature after the enzymolysis and drop, then wash, collect the cell.
In the above technical scheme, in step (5), after the cell culture on the set of shaped scaffold body is completed, when the cell is required to be subcultured, a brand new shaped scaffold body is taken, the edge of the brand new shaped scaffold body is spliced with the shaped scaffold body with the cultured cell, the brand new shaped scaffold body is spliced through the projection and the groove, and the brand new shaped scaffold body is immersed in the culture medium, so that the cell on the original shaped scaffold body can migrate, proliferate or differentiate to the new shaped scaffold body, and the subculture is completed.
In the technical scheme, the preforming support body is integrally formed by 3D printing of a metal material or a non-metal material; when the non-metal material is adopted, a photoinitiator or a thickening agent is added into the non-metal material according to different types of digital 3D printers.
Because of the application of the technical scheme, compared with the prior art, the utility model has the following advantages:
1. the utility model discloses in set up the cell culture hole on the support body, and the support body adopts 3D printing technique to print the shaping with support body an organic whole, and the support body that 3D printed provides a three-dimensional growth environment that more is close to the internal for cell culture, has increased the secretion of extracellular matrix, has strengthened the contact between the cell for the biological characteristic of cell can be fully expressed, does benefit to the biological activity who arouses the cell more, effectively improves the cultivation effect of cell;
2. the utility model adopts the support body with the 3D structure, compared with the prior plane culture proliferation structure adopting the 2D structure, the support body with the 3D structure can provide a larger culture area for adherent cells to be attached and proliferated, thereby effectively improving the culture efficiency;
3. the utility model discloses well support body possesses lug and recess for splice with adjacent support body, constitutes a mosaic structure, can splice a new support body after cell grows over a support body like this to reach the migration, the hyperplasia, the differentiation function of cell, and mosaic structure, can carry out the concatenation of expansion formula, can realize cell proliferation in batches, has not only stopped the pollution that causes because of manual operation in the subculture process, and can increase culture efficiency, can also realize large batch culture;
4. the utility model adopts inert material or material with biological activity, which can not generate negative effect on cell proliferation;
5. the utility model discloses the surface of well support body has the coating, adopts the coating that has increase cell adhesion and cell activity function, can increase the culture quality and the cultivation efficiency of cell like this.
Drawings
Fig. 1 is a schematic structural view of a bracket body according to a first embodiment of the present invention;
fig. 2 is a schematic perspective view of a bracket body according to a first embodiment of the present invention;
FIG. 3 is a schematic view of a structure of a plurality of sets of stent bodies in mutual overlapping fit in a first embodiment of the present invention (cell culture wells are not shown).
Wherein: 1. a stent body; 2. a cell culture well; 3. a bump; 4. and (4) a groove.
Detailed Description
The invention will be further described with reference to the following drawings and examples:
the first embodiment is as follows: referring to fig. 1-3, a cell culture 3D printing support comprises a support body 1, the support body is a 3D printing integrated structure, a plurality of groups of cell culture holes 2 are arranged on the support body, and two ends of each cell culture hole are respectively communicated with the outer wall of the support body.
In the embodiment, the support body is integrally formed by 3D printing and is of a 3D structure, when the support body is used for cell culture, the support body is placed in a culture apparatus, cells and culture media to be cultured are placed in the support body, the cells can be cultured on the support body, and in the culture process, due to the existence of cell culture holes, the support body can provide more culture areas which are hundreds of times larger than the surface area of a conventional culture dish, the limitation of the traditional physical space is solved, the adhesion and proliferation of the cells are facilitated, the culture efficiency is effectively improved, meanwhile, the support body is of the 3D structure, a three-dimensional growth environment which is closer to the in-vivo is provided for the cell culture, the secretion of extracellular matrix is increased, the contact among the cells is enhanced, the bioactivity of the cells is more favorably excited, the biological characteristics of the cells can be fully expressed, and the culture quality of the cells can be improved.
The existence of cell culture hole, give a space of cell culture, wherein, the aperture size of cell culture hole can be unanimous, also can be inconsistent, and the cross-section in cell culture hole can be for circular, ellipse, polygon etc. designs according to the kind and the characteristics that need cultivate the cell, then 3D prints integrated into one piece can, as long as the high-quality multiplication that the required cell of cultivating can be quick of being convenient for is cultivateed can. Simultaneously, the cell culture hole can be the linear type structure, also can be from one end arc structure down to the other end, and the structure is unrestricted, as long as the both ends in cell culture hole respectively with the outer wall intercommunication of support body can, give cell culture's passageway and space like this.
Meanwhile, in this embodiment, the pore size of the cell culture pores of the scaffold body for culturing the same cell can be adjusted, thereby realizing multiple functions of migration, proliferation, differentiation, and the like of the same cell.
Referring to fig. 1 to 3, the outer wall of the bracket body is provided with a projection 3 and a groove 4, and the projection can be clamped in the groove of the adjacent bracket body.
In this embodiment, through the setting of lug and recess, the lug can be gone into in the recess of adjacent support body, the support body is when using like this, adopt the concatenation form, can be after a set of support body is overgrown to the cell, splice a brand-new support body again, thereby reach cell proliferation, the migration, functions such as differentiation, can carry out the concatenation of expansion formula, can realize the cell and proliferate in batches greatly, the pollution of subculture in-process has not only been stopped, and can increase culture efficiency, can also realize big batch, large-scale cultivation.
Referring to fig. 1 to 3, two sets of protrusions and two sets of grooves are respectively disposed on the outer wall of the bracket body, the two sets of protrusions are respectively disposed on two adjacent side walls of the bracket body, the two sets of grooves are respectively disposed on the other two adjacent side walls of the bracket body, and each set of grooves is disposed on the opposite side of one set of protrusions.
In this embodiment, take two sets of lugs to set up respectively on support body right flank and leading flank as an example, then two sets of recesses set up respectively on support body left flank and trailing flank, around a set of support body, can both carry out the concatenation of support body like this, also the support body can be towards the four directions concatenation exactly, the structural optimization of concatenation expansion can splice the expansion from each position, the occupation of reduction space as far as reduces the restriction of expansion.
The length of the bracket body is 5-80 mm, the width of the bracket body is 5-80 mm, and the height of the bracket body is 3-30 mm; the aperture of the bacteria culture hole is 100-800 microns, and the porosity of the bracket body is 50-90%.
In this embodiment, the size of the stent body is preferably 10 mm in both length and width, and the height is selected to be 5 mm to 10 mm.
The stent body is provided with a coating, and the coating is arranged on the outer wall of the stent body and the inner wall of the cell culture hole.
The coating is one or more of collagen, methacrylated collagen, poly-D-lysine, silk fibroin, cellular fibronectin, laminin, vitronectin, gelatin and methacrylated gelatin.
In this embodiment, the coating has the functions of increasing cell adhesion and increasing the biological activity of the stent, so that the cell can better grow on the stent body in the cell culture process, the culture efficiency is improved, and the coating is a material with biological activity and does not have negative influence on cell proliferation. Simultaneously, the coating is degradable material, after the support body has been good to the cell culture, when needing to collect, can adopt specific enzyme to carry out quick enzymolysis with the coating for the cell that cultivates drops naturally, is washed and carries out quick collection, and subsequent collection of this kind of operation is convenient for, and the enzymolysis also can not produce other influences to the cell in addition.
The stent body is made of a metal material or a non-metal material, and the metal material is one or more of titanium, titanium alloy, tantalum metal, stainless steel and cobalt-based alloy; the non-metallic material is one or more of polyethylene glycol and a polymer thereof, polyethylene glycol diacrylate, collagen, methacrylic acid esterified collagen, gelatin, methacrylic acid esterified gelatin, alumina, zirconia, polyetheretherketone, hydroxyapatite, tricalcium phosphate and silicate nano clay.
In this embodiment, when the stent body is made of a metal material or a non-metal material, an inert material or a material having biological activity is used, which does not adversely affect cell proliferation. Furthermore, with biologically active materials, e.g., collagen, can be enzymatically hydrolyzed by the addition of specific enzymes (e.g., collagenase), which facilitates the collection of subsequently cultured cells.
In this embodiment, the stent body is also suitable for various cells with an adherence function, such as various cancer cells, stem cells and immune cells, including but not limited to various cells with an adherence function, such as various solid tumor cells, mesenchymal stem cells, IPS cells, breast cells, fibroblasts, nerve cells, endothelial cells, keratin cells, T cells, NK cells, and the like.
In order to achieve the above purpose, the utility model discloses a cell culture method of cell culture 3D printing support, its step is:
(1) designing a support body through three-dimensional software to obtain a digital support model, wherein the digital support model has the same shape parameters as the designed support;
(2) integrally printing and molding the digital support model obtained in the step (1) by using a digital 3D printer to obtain a preformed support body;
(3) cleaning and drying the preformed support body;
(4) sterilizing the preformed support body in the step (3) to obtain a formed support body; wherein, gamma irradiation sterilization or electron beam irradiation sterilization can be adopted;
(5) placing the formed bracket body into a culture dish, a culture plate or a culture bottle, and dripping cells and a culture medium to be amplified on the formed bracket body by using a dropper to ensure that the formed bracket body is fully immersed by the culture medium;
(6) putting a culture dish, a culture plate or a culture bottle which is provided with the molded bracket body and the cells into a carbon dioxide incubator for culturing and periodically replacing a culture medium; when the cells are cultured, the outer wall of the cell forming bracket body and the cell culture holes are subjected to enrichment culture, and the culture space and the area are large.
Meanwhile, when Sub-culture is required, a brand new forming bracket body is taken, the edge of the brand new forming bracket body is spliced with the original forming bracket through a bump and a groove, and the brand new forming bracket body is immersed in a culture medium, so that the migration function of cells is quickly realized;
(7) and (4) washing after the cell culture achieves the use purpose, and collecting the cells.
Before sterilizing the preformed support body, immersing the preformed support body in the solution of the coating material, standing for 4-24 hours, taking out the preformed support body, drying the preformed support body to finish coating, and after finishing coating, performing the sterilization treatment of the step (4).
In this embodiment, the surface of the stent body may not be provided with a coating layer, or may be provided with a coating layer, and if the coating layer is required to be provided, the coating operation may be performed before sterilization according to the above steps.
Aiming at the forming support body with the coating, after the cell culture achieves the use purpose, specific enzyme is added into the forming support body or the forming support body with the coating, so that the coating or the forming support body is subjected to enzymolysis by the specific enzyme, the cells naturally fall off after the coating or the forming support body is subjected to enzymolysis, then the washing is carried out, and the cultured cells are collected. For example, collagen protease is added into a coating of a collagen material, cells naturally fall off after the coating is subjected to enzymolysis, and then washing is carried out, so that the cultured cells can be collected.
In the step (6), after the cell culture on the set of molded support bodies is completed, when the cell is required to be subcultured, a brand new molded support body is taken, the edge of the brand new molded support body is spliced with the molded support body of the cultured cell, the bump and the groove are spliced, and the brand new molded support body is immersed in the culture medium, so that the cell on the original molded support body can migrate, proliferate or differentiate to the new molded support body, and the subculture is completed.
Meanwhile, in this embodiment, when the cells are cultured, the cells may be cultured by using an undegradable material as the stent body, or a material with a particularly slow degradation rate (which takes several months or more to degrade) as the stent body. When cultured cells need to be collected subsequently, materials capable of being degraded quickly are adopted, or the surface of the bracket body is provided with a coating capable of being degraded quickly, then the bracket body is spliced with the previous bracket body through a bump and a groove, so that the cultured cells can be migrated to a new bracket body, then an enzymolysis mode is adopted, after the coating or the bracket body is subjected to enzymolysis, the cultured cells naturally fall off, and then the bracket body is washed and collected.
The preforming support body is integrally formed by 3D printing of a metal material or a non-metal material; when the non-metal material is adopted, the photoinitiator or the viscous agent is added into the non-metal material according to different types of the digital 3D printer.
In this embodiment, the stent body is made of a metal material or a non-metal material. When the metal material is adopted, a selective laser melting 3D printing technology or a selective electron beam melting 3D printing technology and a laser cladding 3D printing technology are adopted.
When the non-metal material is adopted, an extrusion type precipitation molding 3D printing technology or a stereolithography 3D printing technology is adopted, wherein a thickening agent such as methyl cellulose is required to be added when the non-metal material cannot form a viscous state, so that the non-metal solution is changed into a gel state, and the extrusion of the non-metal material is facilitated for 3D printing molding; the stereolithography 3D printing technology comprises a surface light source 3D printing technology and a point light source 3D printing technology, when the stereolithography 3D printing technology is adopted, photoinitiators of 0.2% -2%w/v are arranged in a non-metal material, and the photoinitiators comprise one or more of Irgacure2959, phenyl (2,4,6-trimethylformyl), lithium phosphate, TPO, procyanidine and ethyl pyruvate.
Claims (6)
1. The utility model provides a cell culture 3D prints support which characterized in that: including the support body, the support body prints the integrated into one piece structure for 3D, be equipped with multiunit cell culture hole on the support body, the both ends in cell culture hole respectively with the outer wall of support body is linked together.
2. The cell culture 3D printing support of claim 1, wherein: the outer wall of the support body is provided with a convex block and a groove, and the convex block can be clamped in the groove of the adjacent support body.
3. The cell culture 3D printing support of claim 2, wherein: the outer wall of the support body is respectively provided with two groups of convex blocks and two groups of grooves, the two groups of convex blocks are respectively arranged on two adjacent side walls of the support body, the two groups of grooves are respectively arranged on the other two adjacent side walls of the support body, and each group of grooves is arranged on the opposite sides of one group of convex blocks.
4. The cell culture 3D printing support of claim 1, wherein: the length of the bracket body is 5-80 mm, the width of the bracket body is 5-80 mm, and the height of the bracket body is 3-30 mm; the aperture of the cell culture hole is 100-800 microns.
5. The cell culture 3D printing support of claim 1, wherein: the porosity of the stent body is 50% -90%.
6. The cell culture 3D printing support of claim 1, wherein: the bracket body is provided with a coating, and the coating is arranged on the outer wall of the bracket body and the inner wall of the cell culture hole.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202222024383.9U CN218089628U (en) | 2022-08-02 | 2022-08-02 | Cell culture 3D prints support |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202222024383.9U CN218089628U (en) | 2022-08-02 | 2022-08-02 | Cell culture 3D prints support |
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| CN218089628U true CN218089628U (en) | 2022-12-20 |
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| CN202222024383.9U Active CN218089628U (en) | 2022-08-02 | 2022-08-02 | Cell culture 3D prints support |
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- 2022-08-02 CN CN202222024383.9U patent/CN218089628U/en active Active
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