CULTURE DEVICE FOR TISSUE CELL SUSPENSION

Abstract

The present invention relates to a culture device for tissue cell suspension. The culture device includes a tissue cell culture body. The tissue cell culture body is a porous material. The porous material is formed by cavities classified into different levels according to the pore size of material and cavity walls surrounding to form the cavities. A lower level of small cavities is provided to surround the cavity wall which forms the upper level of large cavity. Cavities of each level are in communication with each other, and cavities between respective levels are also in communication with each other. The culture device further includes a swirler device provided therein. Such culture device can particularly facilitate normal and unrestricted growth of the suspension of cells in three-dimensional space, obstructing the formation of over-dense cell region or nutrient-rich region during the cell culturing.

Claims

1. A culture device for tissue cell suspension, comprising: a tissue cell culture body; wherein the tissue cell culture body is a porous material; the porous material is formed by a plurality of cavities classified into different levels according to a pore size of the porous material and a plurality of cavity walls surrounding to form the plurality of cavities; a lower level of small cavities is provided to surround the plurality of cavity walls which forms an upper level of large cavities; the plurality of cavities of a same level of the lower level and the upper level are in communication with each other; the plurality of cavities among different levels of the lower level and the upper level are also in communication with each other; the culture device further includes a swirler device disposed therein.

2. The culture device for tissue cell suspension of claim 1, wherein the plurality of levels comprise at least two levels; a next level of the plurality of the small cavities are provided on the plurality of cavity walls forming the upper level of the large cavities; or the lower level comprises a plurality of subordinate levels, wherein the subordinate levels of the plurality of small cavities are provided on the plurality of cavity walls forming the upper level of large cavities; or any combination of the subordinate level of the plurality of small cavities are provided on the cavity wall forming the upper level of large cavities.

3. The culture device for tissue cell suspension of claim 1, wherein the plurality of cavities are classified into three levels; wherein a next level of small cavities is provided to surround the cavity wall forming the upper level of large the cavities; or two lower levels of small cavities are provided to surround the cavity wall forming the upper level of large cavities.

4. The culture device for tissue cell suspension of claim 1, wherein regarding the porous material as the tissue cell culture body, a pore size of the lower level of cavities is nanoscale, and a pore size of other levels of cavities is at least more than an average diameter of the culture cells.

5. The culture device for tissue cell suspension of claim 1, wherein regarding the porous material as the tissue cell culture body, a pore size of the lower level of cavities is nanoscale, and a pore size of other levels of cavities is at least more than twice of an average diameter of the culture cells.

6. The culture device for tissue cell suspension of claim 3, wherein regarding the porous material as the tissue cell culture body, a next level of small cavities are provided to surround the cavity wall forming an upper level of large cavities; the pore size of the lowest level of cavities is nanoscale; a pore size of the cavity which is one level upper than the pore size of the lowest level of cavities is at least more than twice of an average diameter of the culture cells; a pore size of the cavity which is two levels upper than the pore size of the lowest level of small cavities is at least more than four times of an average diameter of the culture cells.

7. The culture device for tissue cell suspension of claim 1, wherein the culture device further includes a culture dish accommodating the tissue cell culture body; the tissue cell culture body is fixed in the culture dish; the swirler device is provided below the tissue cell culture body.

8. The culture device for tissue cell suspension of claim 7, wherein the swirler device includes multiple groups of air transmission pipes supported by the holder; each of the multiple groups of air transmission pipes is provided with an outlet pipe and an intake pipe; the intake pipe is connected to the control valve outside the culture dish; the control valve is also connected to a main intake pipe.

9. The culture device for tissue cell suspension of claim 8, wherein the air transmission pipe is annular; the annular air transmission pipe is provided with an outlet pipe; a projection of an axial line of the outlet pipe in the horizontal plane is tangential with respect to a projection of an axial line of the annular air transmission pipe in the horizontal plane.

10. The culture device for tissue cell suspension of claim 8, wherein the multiple groups of air transmission pipes are even groups of air transmission pipes; on the projection of the horizontal plane, a direction of one group of outlet pipes is the same as a clockwise tangential direction of the axial line of the air transmission pipe, while a direction of the other group of outlet pipes is the same as an anti-clockwise tangential direction of the axial line of the air transmission pipe.

11. The culture device for tissue cell suspension of claim 8, wherein an angle of the outlet pipe with respect to the horizontal plane is less than or equal to 45.

12. The culture device for tissue cell suspension of claim 8, wherein an exit of the outlet pipe is provided with a screen mesh or a membrane with pores.

13. The culture device for tissue cell suspension of claim 2, wherein regarding the porous material as the tissue cell culture body, a pore size of the lower level of cavities is nanoscale, and a pore size of other levels of cavities is at least more than an average diameter of the culture cells.

14. The culture device for tissue cell suspension of claim 2, wherein regarding the porous material as the tissue cell culture body, a pore size of the lower level of cavities is nanoscale, and a pore size of other levels of cavities is at least more than twice of an average diameter of the culture cells.

15. The culture device for tissue cell suspension of claim 2, wherein the culture device further includes a culture dish accommodating the tissue cell culture body; the tissue cell culture body is fixed in the culture dish; the swirler device is provided below the tissue cell culture body.

16. The culture device for tissue cell suspension of claim 9 wherein the multiple groups of air transmission pipes are even groups of air transmission pipes; on the projection of the horizontal plane, a direction of one group of outlet pipes is the same as a clockwise tangential direction of the axial line of the air transmission pipe, while a direction of the other group of outlet pipes is the same as an anti-clockwise tangential direction of the axial line of the air transmission pipe.

17. The culture device for tissue cell suspension of claim 9, wherein an angle of the outlet pipe with respect to the horizontal plane is less than or equal to 45.

18. The culture device for tissue cell suspension of claim 9, wherein an exit of the outlet pipe is provided with a screen mesh or a membrane with pores.

19. The culture device for tissue cell suspension of claim 3, wherein regarding the porous material as the tissue cell culture body, a pore size of the lower level of cavities is nanoscale, and a pore size of other levels of cavities is at least more than an average diameter of the culture cells.

20. The culture device for tissue cell suspension of claim 3, wherein regarding the porous material as the tissue cell culture body, a pore size of the lower level of cavities is nanoscale, and a pore size of other levels of cavities is at least more than twice of an average diameter of the culture cells.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Hereinafter, with reference to drawings and embodiments, the present invention is further illustrated.

[0033] FIG. 1 is a schematic diagram of the tissue cell culture device of the present invention.

[0034] FIG. 2 is a sectional diagram taken along A-A in FIG. 1.

[0035] FIG. 3 is a partially enlarged view of the structure of the tissue cell culture body in FIG. 1.

[0036] FIG. 4 is a partially enlarged diagram of the cavity wall of the tissue cell culture body in FIG. 3.

[0037] FIG. 5 is a sectional diagram taken along B-B in FIG. 4.

[0038] FIG. 6 is a partial view taken in the direction of C in FIG. 2.

[0039] FIG. 7 is a partially enlarged view taken in the direction of D in FIG. 6.

DETAILED DESCRIPTION OF THE INVENTION

[0040] Hereinafter, with reference to drawings, embodiments of the present invention are described. With the technical solution of the present invention as premises, embodiments provide detailed embodiments and specific operation procedure. However, the protective scope of the present invention is not limited to the following embodiments.

[0041] In the drawings; 1 is the culture dish; 2 is the cell culture body; 3 is the main intake pipe; 4 is the holder supporting the air transmission; 5 is the control valve; 6 is the first intake pipe; 7 is the second intake pipe; 8 is the second air transmission pipe; 9 is the first air transmission pipe; 10 is the second outlet pipe; 11 is the first outlet pipe; 12 is the cavity of the tissue cell culture body; 13 is the cavity wall of the cavity; 14 is the cavity on cavity wall 13; 15 is the cavity wall of cavity 14; 16 is a screen mesh or a membrane with pores installed on an exit of the first outlet pipe 11 or the second outlet pipe 10.

[0042] Embodiments of the present invention are provided in detail as below:

Embodiment 1

[0043] Referring to the drawings, a culture device for tissue cell suspension is provided. The culture device includes a tissue cell culture body 2. The tissue cell culture body 2 is a porous tantalum material. The porous tantalum material is formed by cavities classified into different levels according to the pore size of the material and cavity walls surrounding to form the cavities. A lower level of small cavity is provided on the cavity wall which are enclosed to form the upper level of large cavity. Cavities of each level are connected with each other, and cavities between respective levels are also connected with each other. The culture device further includes a swirler device provided therein.

[0044] Particularly in the embodiment, the porous tantalum material includes three levels of cavity, wherein cavity wall 13 of a second upper level of large cavities 12 (hereinafter, referred as the first level of cavities), that are uniformly distributed and are connected with each other, is provided with a first upper level of large cavities 14 (hereinafter, referred as the second level of cavities) that are uniformly distributed and are connected with each other. Cavity wall 15 of the second level of cavities 14 is provided with a lower level of small cavities (hereinafter, referred as the third level of cavities) that are uniformly distributed and are connected with each other. Cavities between respective levels are also connected with each other. The pore size of the first level of cavities 12 is 160 m-650 m. The pore size of the second level of cavities 14 is 60 m-90 m. The pore size of the third level of cavities is 400 nm-700 nm.

[0045] The preparation method of the porous tantalum material is as below:

[0046] (1) Material Preparation

[0047] Tantalum powder with a grain size of 1 m-10 m is used as the raw material. Urea with a grain size of 450 nm-780 nm is used as the pore-forming agent of the lowest level of pores. Polystyrene with a grain size of 450 nm-780 nm is used as the adhesive. The slurry is prepared based on a volume ratio of 1:2:1:7.5 based on tantalumpowder:urea:polystyrene:distilled water.

[0048] Polyester foam with a size of 100 m-200 m is used. The slurry is filled therein uniformly with the foam impregnation method to form a green body. The green body is dried. Next, the green body is crushed, so as to obtain mixed grains with a grain size of 651 m-100 m, including the raw material, the pore-forming agent, and the polyester foam.

[0049] (2) The mixed grains and ethyl cellulose with a grain size of 65 m-100 m are mixed uniformly based on a volume ratio of 2:1. The mixture is poured uniformly into the polyester foam with a strut diameter of 2001 m-720 m and a size of 400 m-600 m, which is three-dimensional transfixion. Next, the polyester foam is disposed into a closed mold to be pressed into a compact green body.

[0050] (3) The compact green body is sintered in vacuum. The sintered green body is subjected to a regular follow-up heat treatment based on tantalum material process, so as to obtain the porous tantalum with three levels of pores.

[0051] Further referring to the drawings, it can be seen that the tissue cell culture device of the present embodiment further includes culture dish 1 accommodating the tissue cell culture body. The tissue cell culture body 2 is fixed in the culture dish 1. The swirler device is provided below the tissue cell culture body.

[0052] The swirler device includes two groups of air transmission pipes supported by holder 4. The air transmission pipes are annular. That is, in the Figure, the first air transmission pipe 9 and the second air transmission pipe 8 both are supported by holder 4. The first air transmission pipe 9 and the second air transmission pipe 8 are evenly provided with the first outlet pipe 11 and the second outlet pipe 12, that are connected with the first air transmission pipe 9 and the second air transmission pipe 8 respectively. The projection of the axial line of the first outlet pipe 11 and that of the second outlet pipe 12 in the horizontal plane are respectively tangential with respect to the projection of the axial line of the first air transmission pipe 9 and that of the second air transmission pipe 8 in the horizontal plane. The first intake pipe 6 and the second intake pipe 7 respectively are connected with the first air transmission pipe 9 and the second air transmission pipe 8 go through the culture dish 1 to be connected to control valve 5. Main intake pipe 3 is connected to control valve 5. Control valve 5 can control the air to go through either the first intake pipe 6 or the second intake pipe 7 at the same time. In the projection of the horizontal plane, the direction of the first outlet pipe 11 is the same as the clockwise tangential direction of the axial line of the first air transmission pipe ring 9, while the direction of the second outlet pipe 12 is the same as the anti-clockwise tangential direction of the axial line of the second air transmission pipe ring 8.

[0053] 1 and 2 that are angles of the first outlet pipe 11 and the second outlet pipe 12 with respect to the horizontal plane are 30.

[0054] Exits of the first outlet pipe 11 and the second outlet pipe 12 are provided with screen mesh 16. The average size of screen mesh 16 is 0.5 mm.

[0055] During the cell culturing, DMEM (Dulbecco's Modified Eagle Medium) culture solution is used. S180 sarcoma cells as the culture cells are disposed into tissue cell culture body 2. Next, tissue cell culture body 2 is disposed into culture dish 1. The first intake pipe 6 and the second intake pipe 7 alternately take in air respectively, 3 minutes each time.

[0056] The culture results show that the body of the cultured cell is fill, and the growth state is good.

Embodiment 2

[0057] Cell culture body 2 of the tissue cell culture device of the present embodiment uses porous silicon dioxide material, which has two levels of cavities. Cavity wall 13 of an upper level of large cavities 12, that are uniformly distributed and are connected with each other, is provided with a lower level of small cavities 14 that are uniformly distributed and are connected with each. Two levels of cavities are also connected with each other. The pore size of the upper level of large cavities 12 is 20 m-80 m. The pore size of the lower level of small cavity 14 is 100 nm-300 nm. 1 and 2 that are angles of the first outlet pipe 11 and the second outlet pipe 12 with respect to the horizontal plane are 15. Exits of the first outlet pipe 11 and the second outlet pipe 12 are provided with a membrane with pores 16, the average size of which is 0.3 mm.

[0058] The preparation method of such kind of porous silicon dioxide material is as below:

[0059] (1) Material Preparation

[0060] Silicon dioxide powder with a size of 1 m-10 m is used. Urea with the grain size of 130 nm-350 nm is used as the pore-forming agent of the lowest level of pores. The silicon dioxide powder and the urea are mixed uniformly. Starch with a size of 130 nm-350 nm is used as the adhesive. The slurry is prepared based on a volume ratio of 1:1.5:1:7 based on silicon dioxide powder:urea:starch:distilled water.

[0061] The slurry is filled uniformly into polyester foam with a strut diameter of 25 m-90 m with the foam impregnation method, so as to form a green body. The green body is dried. Next, the green body is crushed, so as to obtain mixed grains with a grain size of 25 m-90 m, including the silicon dioxide powder, the pore-forming agent, and polyester foam.

[0062] (2) The mixed grains and methyl cellulose with a grain size of 25 m-90 m are mixed uniformly based on a volume ratio of 2:1. Next, the mixture is disposed into a closed mold to be pressed into the compact green body.

[0063] (3) The compact green body is sintered in vacuum. The sintered green body is subjected to regular follow-up heat treatment based on silicon dioxide process, so as to obtain the porous silicon dioxide with two levels of pores.

[0064] Remaining steps are the same as those in Embodiment 1.

[0065] During the cell culturing, BME (Basal Medium Eagle) culture solution is used. Lymph cells as the culture cells are disposed into tissue cell culture body 2. Next, tissue cell culture body 2 is disposed into culture dish 1. The first intake pipe 6 and the second intake pipe 7 alternately take in air respectively, each time 5 minutes.

[0066] The culture results show that the body of the cultured cell is full, and the growth state is good.

Embodiment 3

[0067] Cell culture body 2 of the tissue cell culture device of the present embodiment uses porous titanium material, which includes three levels of cavities. Cavity wall 13 of a second upper level of large cavities 12, (hereinafter, referred as the first level of cavity) that are uniformly distributed and are connected with each other, is provided with a first upper level of large cavities 14 (hereinafter, referred as the second level of cavity) that are uniformly distributed and are connected with each other. Cavity wall 15 of the upper level of large cavities is provided with a lower level of small cavities (hereinafter, referred as the third level of cavities), that are uniformly distributed and are connected with each other. Cavities between respective levels are also connected with each other. The pore size of the first level of cavities 12 is 300 m-700 m. The pore size of the second level of cavities 14 is 60 m-95 m. The pore size of the third level of cavities is 400 nm-700 nm. 1 and 2 that are angles of the first outlet pipe 11 and the second outlet pipe 12 with respect to the horizontal plane are 45. Exits of the first outlet pipe 11 and the second outlet pipe 12 are provided with a membrane with pores 16, the average size of which is 0.7 mm.

[0068] The preparation method of the porous titanium material is as below:

[0069] (1) Material Preparation

[0070] Titanium powder grain with a size of 1 m-10 m is used as the raw material. The urea with a grain size of 430 nm-750 nm is used as the pore-forming agent of the lowest level of pores. Polystyrene with a grain size of 430 nm-750 nm is used as the adhesive. The slurry is prepared based on a volume ratio of 1:2:1:7.5 based on titanium powder:urea:polystyrene:distilled water.

[0071] Polyester foam with a size of 100 m-200 m is used, and the slurry is filled therein uniformly with the foam impregnation method to form a green body. The green body is dried. Next, the green body is crushed, so as to obtain mixed grains with a grain size of 65 m-105 m, including the raw material, the pore-forming agent, and the polyester foam.

[0072] (2) The mixed grains and ethyl cellulose with a grain size of 65 m-105 m are mixed uniformly based on a volume ratio of 2:1. The mixture is uniformly into the polyester foam strut diameter of 350 m-780 m and a size of 400 m-600 m, which is three-dimensional communicating. Next, the polyester foam is disposed into a closed mold to be pressed into a compact green body.

[0073] (3) The compact green body is sintered in vacuum. The sintered green body is subjected to regular follow-up heat treatment based on titanium material process, so as to obtain porous titanium with three levels of pores.

[0074] Remaining steps are the same as those in Embodiment 1.

[0075] During the cell culturing, MEM (Minimum Essential Medium) culture solution is used. Culture cells K562 are disposed into tissue cell culture body 2. Next, tissue cell culture body 2 is disposed into culture dish 1. The first intake pipe 6 and the second intake pipe 7 alternately take in air respectively, 4 minutes each time.

[0076] The culture results show that the body of the cultured cell is full, and the growth state is good.