CN116410862A - Large specific surface area on-line cell culture device - Google Patents
Large specific surface area on-line cell culture device Download PDFInfo
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- CN116410862A CN116410862A CN202310199104.9A CN202310199104A CN116410862A CN 116410862 A CN116410862 A CN 116410862A CN 202310199104 A CN202310199104 A CN 202310199104A CN 116410862 A CN116410862 A CN 116410862A
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS 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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/20—Material Coatings
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- C12M—APPARATUS 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
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/10—Hollow fibers or tubes
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS 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
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/16—Hollow fibers
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Abstract
The invention provides an online cell culture device with large specific surface area, which is characterized in that yarns with hairiness are arranged on the surfaces of hollow fibers of the culture device, the performances such as the outer diameter, the length and the porosity of the hollow fibers are regulated and controlled, and meanwhile, the linear density, the screw pitch and the performances of the hairiness on the surfaces of the yarns are regulated, so that a flow channel is provided for culture fluid by using the hollow fibers, the hollow fibers are connected by using the hairiness of the yarns, a bracket is provided for reproduction of cells, and the culture cell density of the cell culture device is effectively enlarged; the preparation process comprises winding the yarn on the surface of the hollow fiber, and bonding to form a uniform whole between the yarn and the hollow fiber, so that the stability of the cell culture device in the cell culture process is ensured. Through the mode, the online cell culture device with the large specific surface area is simple in preparation method, low in cost and good in stability.
Description
Technical Field
The invention relates to the technical field of cell culture, in particular to an online cell culture device with a large specific surface area.
Background
Cell culture refers to a technology for simulating in-vivo environment in vitro to enable cells to survive, grow, reproduce and maintain main structures and functions, and can directly observe morphological structures and vital activities of living cells and easily provide a large number of experimental objects with similar biological properties. Cell culture has become a new approach adopted in the field of biological and medical research. The cells prepared by the culture method can be directly used for the circulatory system of human body and treated by the cells. The principle of the method is to provide necessary conditions for in vitro cell culture, including space conditions, liquid conditions, gas conditions, temperature conditions, clean conditions and nutrition conditions for cell growth, in particular to conditions such as a cell culture medium, a sterile environment, specific gas or liquid concentration and the like, pH value of a culture solution, environmental temperature and the like. At present, the culture method is divided into adherent cell culture and suspension cell culture according to the cell growth culture mode, and the adherent culture has the defects of low culture density, large investment, large occupied area, incapability of effectively monitoring cell growth and the like, and the hollow fiber bioreactor technology can provide an effective cell adherent culture unit for adherent animal cells.
In the prior art, the application number is 201710708156.9, the publication date is 2017, 11 and 28, and the patent application document entitled "hollow fiber exchanger and hollow fiber exchange culture system" discloses a hollow fiber exchanger, which comprises an outer wall tube, hollow fiber wires arranged inside the outer wall tube, sealing clamping blocks and end covers arranged at two ends of the outer wall tube, and an interface; the sealing clamping blocks are arranged in the ports at the two ends of the outer wall pipe, fill gaps among all fiber wires and between the fiber wires and the outer wall pipe, are tightly attached to the inner part of the outer wall pipe and play a role in sealing; the end covers are used for sealing two ends of the outer wall pipe, and a fiber yarn inner buffer area is formed between the sealing clamping blocks and the end covers; the inner space of the hollow fiber yarn forms a fiber yarn inner flow path space, and a fiber yarn outer flow path space is formed among the outer wall of the hollow fiber yarn, the inner wall of the outer wall pipe and the sealing clamping block and is communicated with the fiber yarn inner buffer zone. The hollow fiber exchanger can realize large-scale high-density culture of cells, has automatic operation and good stability, but the specific surface area of a culture system of the hollow fiber wires is smaller, which is not beneficial to the adhesion growth of cells.
In the prior art, the application number is 202210714527.5, the publication date is 2022, 8 and 26, and the technical scheme of the hollow fiber membrane with a spiral line structure, the preparation method and the membrane contact reactor is that the hollow fiber membrane with the spiral line structure comprises a hollow fiber base membrane and fiber lines which are wound on the surface of the hollow fiber base membrane in a spiral mode, and the wound fiber lines and the hollow fiber base membrane are adhered to form a continuous whole in a sintering treatment mode. The membrane contact reactor prepared by the hollow fiber membrane can maintain a specific distance between the membranes of the hollow fiber membranes by utilizing the spiral line structure of the outer surface of the hollow fiber base membrane so as to maintain a fluid channel, avoid the mutual extrusion of the membranes, reduce the contact area between the membranes and gas or liquid and realize the smooth proceeding of the mass transfer process. However, the hollow fiber membrane prepared in the above technical scheme has an extremely smooth surface, and when the hollow fiber membrane is used as a cell culture device, the surface area is too small, so that the adhesion, growth and proliferation of cells are not facilitated.
In view of the above, there is a need for an improved large specific surface area in-line cell culture apparatus that solves the above-described problems.
Disclosure of Invention
The invention aims to provide an online cell culture device with a large specific surface area.
In order to achieve the aim of the invention, the invention provides an online cell culture device with large specific surface area, which comprises a columnar shell and a plurality of hollow fibers arranged in parallel in the shell, wherein two opposite ends of the shell are respectively provided with a culture solution inlet and a culture solution outlet, two sides of the shell are provided with a cell perfusion opening and a cell outlet, and yarns with hairiness are wound outside the hollow fibers.
Preferably, the yarn is wound around the outer surface of a single hollow fiber
Preferably, the outer diameter of the hollow fiber is 0.2-1mm; the length of the hollow fiber is 10-80cm; the porosity of the hollow fiber is 40-80%; the pore diameter of the hollow fiber gap is 0.05-10 mu m.
Preferably, the yarn has a hairiness length of 1-6mm; the number of hairiness of the yarn is 500-10000/m; the linear density of the yarn is 10-40tex; the pitch of the yarn shown is 0.3-3mm.
Preferably, the hollow fiber is made of one of polyester, polypropylene, polyacrylonitrile, polysulfone, polyether sulfone and polyvinyl alcohol.
Preferably, the yarn is made of one or more of polyester, polyamide, polypropylene, polyethylene, polyacrylonitrile, polyvinyl alcohol and cellulose fibers.
Preferably, the binder is polyurethane glue, epoxy resin or biological binder.
The beneficial effects of the invention are as follows:
1. the large specific surface area on-line cell culture device provided by the invention is realized by increasing the specific surface area through hairiness, and branched hairiness provides an adsorption area for cells. According to the invention, the yarn with hairiness is fixed on the surface of the hollow fiber, the performances of the outer diameter, the length, the porosity and the like of the hollow fiber are regulated and controlled, and meanwhile, the linear density and the lead of the yarn and the performances of the hairiness on the surface of the yarn are regulated, so that a flow channel is provided for liquid by using the hollow fiber, the hollow fiber is communicated by using the hairiness of the yarn, the culture density of the cell culture device is effectively increased, a good growing environment is provided for cell culture, and the large-scale high-density culture of cells is realized.
2. According to the online cell culture device with large specific surface area, hairiness is formed on yarns, so that the hairiness can be used for providing carriers for cells, the adherent growth of adherent cells is facilitated, meanwhile, the hairiness and the hairiness as well as the parallel yarns can be communicated by utilizing capillary effect among the hairiness, and a channel is provided for cell transportation and mass transfer among hollow fibers; the yarn is wound on the surface of the hollow fiber and then bonded, so that a unified whole is formed between the yarn and the hollow fiber, and the stability of the cell culture device in the cell culture process is ensured.
3. According to the large specific surface area on-line cell culture device provided by the invention, the cell perfusion opening and the cell outlet are arranged on the two sides of the shell, so that a channel is provided for cell injection and cell removal, the cells can be cultured and taken out at any time, and the on-line culture of the cells is realized.
4. The online cell culture device with large specific surface area provided by the invention has the advantages of low yarn price, simplicity, easiness in obtaining and simplicity in winding mode. Through the mode, the online cell culture device with the large specific surface area is simple in preparation method, low in cost and good in stability.
Drawings
FIG. 1 is a schematic diagram of the structure of an in-line cell incubator with a large specific surface area according to the present invention;
FIG. 2 is a schematic view of the yarn of FIG. 1 wrapped with hollow fibers;
FIG. 3 is an interface view of a yarn wrapped with hollow fibers;
the reference numerals are as follows:
1. a housing; 2. hollow fiber; 3. a culture fluid inlet; 4. a culture fluid outlet; 5. a cell perfusion port; 6. a cell outlet; 7. a yarn.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in detail with reference to the accompanying drawings and specific embodiments.
It should be noted that, in order to avoid obscuring the present invention due to unnecessary details, only structures and/or processing steps closely related to aspects of the present invention are shown in the drawings, and other details not greatly related to the present invention are omitted.
In addition, it should be further noted that the terms "comprises," "comprising," or any other variation 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 may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
Referring to fig. 1 to 3, the large specific surface area on-line cell incubator provided by the invention comprises a cylindrical shell 1 and a plurality of hollow fibers 2 which are arranged in the shell 1 in parallel, wherein two opposite ends of the shell 1 are respectively provided with a culture solution inlet 3 and a culture solution outlet 4, two sides of the shell 1 are respectively provided with a cell perfusion opening 5 and a cell outlet 6, the outside of the hollow fibers 2 is wound with yarn 7 with hairiness, and the yarn 7 is wound on the outer surface of a single hollow fiber 2 or is connected with the hollow fibers 2 in a manner of being perpendicular to the hollow fibers 2.
Preferably, the outer diameter of the hollow fiber 2 is 0.2-1mm, the length is 10-80cm, and the porosity is 40-80%; the hollow portion of the hollow fiber 2 has a pore diameter of 0.05 to 10. Mu.m.
Preferably, the hollow fiber 2 is made of one of polyester, polypropylene, polyacrylonitrile, polysulfone, polyethersulfone and polyvinyl alcohol.
Preferably, the yarn 7 has a linear density of 10-40tex and a lead of 0.3-3mm; the hairiness length of the yarn 7 is 1-6mm, and the number of hairiness is 500-10000/m.
Preferably, the material of the yarn 7 is one or more of polyester, polyamide, polypropylene, polyethylene, polyacrylonitrile, polyvinyl alcohol and cotton.
Preferably, the binder is polyurethane glue, epoxy or a biological binder.
The large specific surface area in-line cell culture apparatus of the present invention is further described below with reference to specific examples:
example 1
Referring to fig. 1, the present embodiment provides an online cell incubator with a large specific surface area, which comprises a quadrangular housing 1 and a plurality of hollow fibers 2 made of polyester and arranged in the housing 1 in parallel, wherein two opposite sides of the housing 1 are respectively provided with a culture solution inlet 3 and a culture solution outlet 4, the other side of the housing 1 is provided with a cell perfusion opening 5 and a cell outlet 6, yarns 7 with hairiness are connected to the outer sides of the hollow fibers 2 through an adhesive, the yarns 7 are made of polyester, and the yarns 7 are wound on the outer surfaces of the single hollow fibers 2 and are adhered by using epoxy resin as the adhesive. Specifically, the outer diameter of the hollow fiber 2 was 0.6mm, the length was 30cm, the porosity was 50%, and the pore diameter of the hollow portion of the hollow fiber 2 was 2.5 μm; yarn 7 had a linear density of 20tex and a lead of 0.5mm; the length of hairiness on the yarn 7 is 3mm, and the number of hairiness is 1000 hairiness/m.
The specific surface area of the cell culture vessel measured by BEF test method was 1.50m 2 Per g, the porosity of the cell culture vessel was 50% as measured by mercury intrusion.
Examples 2 to 5
Examples 2 to 5 differ from example 1 in that: the porosity of the hollow fiber is different from that of example 1, and other steps are substantially the same as those of example 1, and are not described here. The setting of the porosity of the hollow fibers of examples 1 to 5 and the performance of the cell culture apparatus produced under the corresponding conditions are shown in table 1, and comparing the number of cells surviving on the surface of the cell culture apparatus produced under different experimental conditions, it was found that the specific surface area of the cell culture apparatus produced gradually increased with the increase of the porosity of the hollow fibers, but the number of cells on the surface of the cell culture apparatus was the largest at 50% porosity, indicating that the cell culture apparatus produced under the conditions was most favorable for cell adhesion and growth.
TABLE 1 setting of porosities of hollow fibers of examples 1 to 5 and Performance of cell incubators produced under corresponding conditions
| Project | Porosity (%) | Specific surface area (m) 2 /g) | Surface cell count |
| Example 1 | 50 | 1.50 | 1.73×10 7 |
| Example 2 | 40 | 1.05 | 9.55×10 6 |
| Example 3 | 60 | 1.43 | 1.53×10 7 |
| Example 4 | 70 | 1.38 | 1.12×10 7 |
| Example 5 | 80 | 1.26 | 1.02×10 7 |
Examples 6 to 10
Examples 6 to 10 differ from example 1 in that: the hairiness length of the hollow fiber is different from that of example 1, and other steps are basically the same as those of example 1, and are not described here again. The hairiness length of the yarns of example 1 and examples 6 to 10 and the performance of the cell culture devices produced under the corresponding conditions are shown in table 2, and comparing the number of cells surviving on the surface of the cell culture devices produced under different experimental conditions, it can be found that the number of surface cells tends to increase and then decrease with increasing hairiness length, and the number of surface cells is the largest when the hairiness length is 3mm.
TABLE 2 hairiness length of yarns of example 1 and examples 6 to 10 and performance of cell culture vessels produced under corresponding conditions
| Project | Hairiness length (mm) | Specific surface area (m) 2 /g) | Surface cell count |
| Example 1 | 3 | 1.50 | 1.73×10 7 |
| Example 6 | 1 | 1.22 | 1.03×10 7 |
| Example 7 | 2 | 1.39 | 1.23×10 7 |
| Example 8 | 4 | 1.37 | 1.17×10 7 |
| Example 9 | 5 | 1.31 | 1.13×10 7 |
| Example 10 | 6 | 1.29 | 1.06×10 7 |
Examples 11 to 15
Examples 11 to 15 differ from example 1 in that: the lead of the hollow fiber 2 is different from that of example 1, and other steps are substantially the same as those of example 1, and will not be described here. The leads of the yarns of example 1 and examples 11 to 15 and the properties of the cell culture devices produced under the corresponding conditions are shown in Table 3, and it was found that the cell adhesion and growth were most advantageous when the leads were 0.5mm, compared with the number of cells surviving on the surface of the cell culture devices produced under the different experimental conditions.
TABLE 3 hairiness length of yarns of example 1 and examples 11 to 15 and performance of cell culture vessels produced under corresponding conditions
| Project | Lead (mm) | Specific surface area (m) 2 /g) | Surface cell count |
| Example 1 | 0.5 | 1.50 | 1.73×10 7 |
| Example 11 | 1 | 1.07 | 9.35×10 6 |
| Example 12 | 1.5 | 0.84 | 7.83×10 6 |
| Example 13 | 2 | 0.65 | 6.10×10 6 |
| Example 14 | 2.5 | 0.47 | 5.11×10 6 |
| Example 15 | 3 | 0.38 | 4.42×10 6 |
Examples 16 to 15
Examples 16 to 15 differ from example 1 in that: the yarn linear density was different from that of example 1, and the other steps were substantially the same as those of example 1, and will not be described again. The linear density of the yarns of example 1 and examples 11 to 15 and the properties of the cell culture devices produced under the corresponding conditions are shown in Table 4, and it was found that the linear density was most favorable for cell adhesion and growth when the number of cells surviving on the surface of the cell culture devices produced under the different experimental conditions was compared with that of the yarns of example 1 and examples 11 to 15.
TABLE 4 Linear densities of the yarns of example 1 and examples 11 to 15 and properties of the cell incubators produced under the corresponding conditions
Comparative examples 1 to 8
Comparative examples 1 to 8 differ from example 1 in that: the porosity of the hollow fiber and the hairiness length, lead and linear density of the yarn were different, and the other steps were substantially the same as in example 1, and will not be described here. The parameter settings of example 1 and comparative examples 1 to 8 and the properties of the cell incubators produced under these conditions are shown in Table 5,
TABLE 5 parameter settings for example 1 and comparative examples 1 to 8 and performance of cell incubators prepared under these conditions
In summary, the large specific surface area on-line cell incubator provided by the invention has the advantages that the yarn with hairiness is arranged on the surface of the hollow fiber of the incubator, the performances such as the outer diameter, the length and the porosity of the hollow fiber are regulated and controlled, and meanwhile, the linear density and the lead of the yarn and the performances of the hairiness on the surface of the yarn are regulated, so that the hollow fiber is utilized to provide a flow channel for liquid, the hollow fiber is utilized to communicate with each other, the culture density of the cell incubator is effectively enlarged, a good growth environment is provided for cell culture, and the large-scale high-density culture of cells is realized. Through the mode, the online cell culture device with the large specific surface area is simple in preparation method, low in cost and good in stability.
The above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications and equivalents may be made thereto without departing from the spirit and scope of the technical solution of the present invention.
Claims (7)
1. The utility model provides a big specific surface area on-line cell culture ware, its characterized in that is including being the casing of column body and parallel arrangement be in a plurality of hollow fiber of the inside of casing, the relative both ends are equipped with culture solution entry and culture solution export respectively on the casing, the casing both sides are equipped with cell perfusion mouth and cell export, hollow fiber's outside winding has the yarn that has the hairiness.
2. The large specific surface area in-line cell incubator of claim 1, wherein the yarn is wrapped around the outer surface of a single hollow fiber.
3. The large specific surface area in-line cell incubator of claim 1, wherein the hollow fiber has an outer diameter of 0.2-1mm; the length of the hollow fiber is 10-80cm; the porosity of the hollow fiber is 40-80%; the pore diameter of the hollow fiber gap is 0.05-10 mu m.
4. The large specific surface area in-line cell incubator of claim 1, wherein the yarn has a hairiness length of 1-6mm; the number of hairiness of the yarn is 500-10000/m; the linear density of the yarn is 10-40tex; the pitch of the yarn shown is 0.3-3mm.
5. The large specific surface area on-line cell incubator of claim 1, wherein the hollow fiber is one of polyester, polypropylene, polyacrylonitrile, polysulfone, polyethersulfone, and polyvinyl alcohol.
6. The large specific surface area on-line cell incubator as recited in claim 1, wherein the yarn is one or more of polyester, polyamide, polypropylene, polyethylene, polyacrylonitrile, polyvinyl alcohol, and cellulose fiber.
7. The large specific surface area on-line cell incubator of claim 1, wherein the adhesive is polyurethane glue, epoxy, or a biological adhesive.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310199104.9A CN116410862A (en) | 2023-03-03 | 2023-03-03 | Large specific surface area on-line cell culture device |
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| CN202310199104.9A CN116410862A (en) | 2023-03-03 | 2023-03-03 | Large specific surface area on-line cell culture device |
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