CN108676716A - A kind of 3D Constituent cells culture carrier and bioreactor - Google Patents
A kind of 3D Constituent cells culture carrier and bioreactor Download PDFInfo
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- CN108676716A CN108676716A CN201810905734.2A CN201810905734A CN108676716A CN 108676716 A CN108676716 A CN 108676716A CN 201810905734 A CN201810905734 A CN 201810905734A CN 108676716 A CN108676716 A CN 108676716A
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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
- C12M23/48—Holding appliances; Racks; Supports
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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
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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
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/02—Stirrer or mobile mixing elements
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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
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/18—Flow directing inserts
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Abstract
The invention discloses a kind of 3D Constituent cells culture carrier and bioreactor, the carriers：Including at least two-layer nonwoven and at least one layer of holder, every layer of frame upper and lower part correspond to be welded and fixed with one layer of non-woven fabrics respectively, the non-woven fabrics is continuous type nonwoven fabric construct or discontinuous form nonwoven fabric construct.The reactor：By tank body, cover, pillar water leg and it is placed in several 3D Constituent cells culture carriers, guiding device and the magnetic stirring apparatus of tank body and forms, tank body is firm by pillar water leg, liquid guide flow is carried out by guiding device and magnetic stirring apparatus, and oxygen is passed for cell mass transfer.The advantages of carrier of the present invention is：Large specific surface area, mass transfer biography oxygen effect is good, and cell density is big；The advantages of reactor of the present invention is：Water conservancy diversion effect is high, and each ingredient mixing efficiency is high, improves the useful load of carrier, convenient for cell amplification culture and technique amplification.
Description
Technical field
The present invention relates to technical field of cell culture, a kind of 3D Constituent cells culture carrier and biology are particularly related to
Reactor
Background technology
Cell culture is an essential process in biotechnology research.Cell type is according to whether attached
In the characteristic grown on support, it is divided into attaching type and floating type.It only relies on and is attached to the cell that support surface could be grown
It is called attaching type cell；It is called floating type cell without attaching the cell grown on the support.Most animals cell has
The custom of adherent growth, therefore when carrying out cultured in vitro, usually with needing to provide support for growth, this support refers to
The cell culture vector that those skilled in the art often say, current domestic cell culture vector common are as 2~11mm of diameter
Particulate vector, porous ball carrier or sheet-like fiber carrier.When carrying out cell culture, the gap between carrier is usually smaller and not
Uniformly, the channel part for be easy to causeing cell solution cycle blocks, and causes mass transfer, biography oxygen and heat transfer effect poor；Secondly, work as cell
When growing into certain density, the support strength of these cell carriers is usually inadequate, be easy to cause carrier in bottom of culture vessel heap
It is poor to will also result in mass transfer, biography oxygen and heat transfer effect for product.Although foreign countries have the sheet for meeting support strength using support frame fine
Cell carrier is tieed up, but there was only planar structure at present, and inactive surface product is relatively more, reduces the specific surface in the useful space
Product.
The bioreactor for carrying out cell culture at present, is broadly divided into fluidized bed aerosol generator and fixed bed biological respinse
Device.Wherein, fixed-bed bioreactor is by loading cell culture vector and guiding device, profit among a fixed container
So that cell culture vector is in suspended motion state with liquid, makes gas is opposite with liquid to should be cell mass transfer using guiding device
Pass the reactor of oxygen；Fluidized bed aerosol generator is a kind of to make solid particle by granular solids layer using gas or liquid
In suspended motion state, the reactor of go forward side by side promoting the circulation of qi solid phase reaction process or liquid and solid phase reaction process；
Existing guiding device is usually made of the guide shell of tablet above and below upper flat plate, lower tablet and connection, is then passed through
Venthole is opened up in upper flat plate and lower tablet to flow back and forth in guide shell by cell culture liquid energy, is embodied as being seated in container
Cell culture vector on cell oxygen or nutrition are provided；But this guiding device is easy that cell carrier and cell is made to put down from upper
The venthole of plate enters guide shell, is easy to block filter screen, and use such guiding device, the cell carrier in reactor
Useful load is fewer, and the attainable density of cell culture institute is smaller, and cell culture process amplification is more difficult, and is also inconvenient for thin
Born of the same parents' culture carrier passes in and out tank interior, and then can not accurately detect the true growth conditions of tank inner cell, can not carry out
Carrier samples, and digestion cannot be carried out to cell in carrier in cell tank and expands culture.Simultaneously when carrying out liquid flowing, filtering is cut
Flow area is small so that fluid exchange efficiency is low, is not easy to being sufficiently mixed for each ingredient.
In addition, existing fixed bed biological respinse also deposits following problem：(1) gas stroke in pipeline is shorter, fully not sharp
With moving toward one another for solution-air, to increase the solubility of gas, solubility of the gas in liquid is relatively poor；(2) cell is carrying
Activity space is small in body, and solution composition homogeneity is poor；(3) nutrition supply effect is poor；(4) state of cell growth is poor,
Low output；(5) agitating paddle can cause cell bigger direct shearing to injure；(5) when being heated to tank body, heating part with it is thin
Born of the same parents have the part being in direct contact, and are easy to keep temperature excessively high and sunburn cell, cause cell death, totally unfavorable to cell culture.
Invention content
The purpose of the present invention is to provide a kind of 3D Constituent cells culture carrier and bioreactors, for solve background
The problems in technology.
To achieve the above object, the technical solution that the present invention takes is：A kind of 3D Constituent cells culture carrier, including 3D is tied
Structure carrier body, the 3D structure carriers ontology include at least two-layer nonwoven and at least one layer of holder, on every layer of holder
Correspondence is welded and fixed with one layer of non-woven fabrics respectively for portion and lower part, described every layer non-woven fabrics for continuous type nonwoven fabric construct or
Discontinuous form nonwoven fabric construct.
Further, when the corresponding every layer of non-woven fabrics of each 3D structure carriers ontology is continuous type nonwoven fabric construct,
Its corresponding every layer of holder be corrugation paper-like or it is latticed in any one；
It is corresponding when the corresponding every layer of non-woven fabrics of each 3D structure carriers ontology is discontinuous form nonwoven fabric construct
Every layer of holder is in latticed, and every layer of non-woven fabrics therein is made of or the non-woven fabric plate of the V-shaped welding of two panels by multiple
The small non-woven fabric plate of setting and V-shaped welding composition at equal intervals.
Further, when the holder is corrugation paper-like, the holder is made of material identical with the non-woven fabrics,
And it is medical grade non-woven fabrics material；
When the holder is latticed, the holder is made of medical grade plastic material.
Further, every layer of nonwoven thickness is 0.1mm~100mm, every layer of backing thickness be 0.1mm~
100mm。
Further, the cross-sectional shape of the 3D structure carriers ontology is any one in circle, ellipse or polygon
Kind；
When the cross-sectional shape of the 3D structure carriers ontology is round, the 3D structure carriers ontology it is a diameter of
1mm~10m is highly 1mm~10m；
When the cross sectional shape of the 3D structure carriers ontology is ellipse, the major axis dimension of the 3D structure carriers ontology
For 1mm~10m, minor axis dimension is 0.9mm~9.9mm, is highly 1mm~10m；
When the cross sectional shape of the 3D structure carriers ontology is polygon, the length of side of the 3D structure carriers ontology is
1mm~10m is highly 1mm~10m.
Further, when the cross-sectional area of the 3D structure carriers ontology is more than 1962.5mm2And 3D structure carrier ontologies
When upper corresponding every layer of non-woven fabrics is continuous type nonwoven fabric construct, several are further opened on the 3D structure carriers ontology
Penetrate through the venthole of lower surface thereon, and a diameter of 1mm~5cm of the venthole.
Several above-mentioned 3D structure carriers ontologies that a kind of bioreactor includes tank body and is placed in the tank body；
Also include the guiding device being vertically installed in the tank body, the guiding device expansion bearing is in the tank body
It is interior；
Also include that bottom and the magnetic stirring apparatus positioned at the guiding device lower part are set in the tank body, it is described
Magnetic stirring apparatus is fixed by welding in the inner bottom wall of the tank body, and the magnetic force generator by being arranged outside tank body
Drive its operation；
Also include the cover being arranged in the top of the tank, the cover is tightly connected with the top of the tank；
Also include the pillar water leg being arranged in the tank body outer bottom, the pillar water leg and the tank body outer bottom
Portion's nested encryptions.
In above-mentioned bioreactor technology scheme, also extended through on the cover carrier probe tube, waste liquid probe tube,
Inlet tube, outlet tube and waste liquid evacuated tube, the carrier probe tube, waste liquid probe tube, inlet tube, outlet tube and waste liquid evacuated tube
The tank interior is stretched into one end, and the other end stretches out the top of the tank.
In above-mentioned bioreactor technology scheme, the guiding device is by double horn above and below top convex baffle, middle part
Mouth guide shell, lower porous baffle and top snorkel form, the upper horn mouth of double horn mouth guide shell above and below the middle part
And lower horn mouth is connect with the top convex baffle and lower porous baffle respectively, and double horn mouth water conservancy diversion above and below the middle part
The lower horn mouth of cylinder is also opposite with the paddle of the magnetic stirring apparatus, and top snorkel one end, which is stretched, is located at middle part up and down
Inside double horn mouth guide shell, the other end is through the cover and stretches out top of the tank.
In above-mentioned bioreactor technology scheme, several filter mesh are further opened on the top convex baffle,
The shape of the top convex baffle is in netted spherical or netted arc surfaced or netted taper；The top convex baffle, in
Portion is linked together between double horn mouth guide shell, lower porous baffle and top snorkel by welding procedure up and down.
In above-mentioned bioreactor technology scheme, it is equipped with and the tank body exterior bottom wall phase at the top of the pillar water leg
Matched groove, the pillar water leg pass through the groove and the tank body outer bottom nested encryptions；It is pressed from both sides in the pillar water
Set is internal to be additionally provided with hollow ring cavity, further respectively has on the left and right sides wall of the pillar water leg and connects with the hollow ring cavity
Logical water inlet pipe and outlet pipe.
The operation principle of bioreactor provided by the invention is as follows：
When work, cell culture fluid enters tank interior by inlet tube, the magnetic stirring apparatus being equipped with by bottom
The effect and immixture for generating flow direction make cell be trained in cell to drive the suspension of cell culture vector slightly up
Support the growth of carrier region；
During cell culture, the monitoring of carrier probe tube and magnocell culture being equipped with by cover pass through
The waste liquid of waste liquid probe tube extraction cell cultivation process, detects nutritional ingredient therein, convenient for replacing or feeding newly thin in time
Born of the same parents' culture solution is ventilated by top snorkel to the cell culture fluid in tank body, to ensure required dissolved oxygen of cell growth etc.；
In the completion of cell culture, the waste liq in tank body is excluded by waste liquid evacuated tube.
Compared with prior art, the beneficial effects of the invention are as follows：
(1) 3D Constituent cells culture carrier provided by the invention has higher specific surface area, can greatly improve cell
Stand density eliminates the hinge assembling process of conventional cell carrier, passes through net grid support or corrugation paper-like branch with gap
Frame forms a 3D structure carrier with non-woven fabrics, and the mass transfer that cell is greatly facilitated passes oxygen, improves the survival rate of cell；
(2) guiding device that bioreactor provided by the invention uses, can greatly promote cell culture fluid in retort
Interior flowing improves the mixing efficiency of each nutritional ingredient of cell so that fluid exchange it is more efficient, mass transfer passes oxygen effect more
It is good, while can also indirectly make the interior more cell culture vectors of filling of the tank body of bioreactor；
(3)) bioreactor provided by the invention is convenient for cell culture convenient for the amplification of the amplification culture and technique of cell
Carrier passes in and out tank interior, thin in culture carrier in the true growth conditions and tank body to accurately detect tank inner cell
The digestion situation of born of the same parents, is also convenient for cell sampling.
(4) bioreactor provided by the invention is by the independent pillar water leg that is equipped in bottom, on the one hand can be with
The support firm effect to tank body is played, external heating or cooling piping facility on the other hand can also be coordinated, is realized to tank body
The purpose for cooling down or heating indirectly, and then it is greatly improved cell survival rate.
Description of the drawings
Fig. 1 is the first embodiment of 3D Constituent cells culture carrier of the present invention
Fig. 2 is the diagrammatic cross-section of the first embodiment；
Second of embodiment of Fig. 3 3D Constituent cells culture carriers of the present invention；
Fig. 4 is the diagrammatic cross-section of second of embodiment；
Fig. 5 is the third embodiment of 3D Constituent cells culture carrier of the present invention；
Fig. 6 is the diagrammatic cross-section of the third embodiment；
Fig. 7 is the 4th kind of embodiment of 3D Constituent cells culture carrier of the present invention；
Fig. 8 is the diagrammatic cross-section of the 4th kind of embodiment；
Fig. 9 is the 5th kind of embodiment of 3D Constituent cells culture carrier of the present invention；
Figure 10 is the 6th kind of embodiment of 3D Constituent cells culture carrier of the present invention；
Figure 11 is the diagrammatic cross-section of the 6th kind of embodiment；
Figure 12 is the 7th kind of embodiment of 3D Constituent cells culture carrier of the present invention；
Figure 13 is the diagrammatic cross-section of the 7th kind of embodiment；
Figure 14 is the 8th kind of embodiment of 3D Constituent cells culture carrier of the present invention；
Figure 15 is the 9th kind of embodiment of 3D Constituent cells culture carrier of the present invention；
Figure 16 is the tenth kind of embodiment of 3D Constituent cells culture carrier of the present invention；
Figure 17 is a kind of the tenth embodiment of 3D Constituent cells culture carrier of the present invention；
Figure 18 is the 12nd kind of embodiment of 3D Constituent cells culture carrier of the present invention；
Figure 19 is the 13rd kind of embodiment of 3D Constituent cells culture carrier of the present invention；
Figure 20 is a kind of embodiment of bioreactor provided by the invention；
Figure 21 is the structural schematic diagram of guiding device in Figure 20；
Figure 22 is the structural schematic diagram of Figure 20 B-C post water legs；
In figure：1,3D structure carriers ontology；1a, non-woven fabrics；1b, holder；1c, venthole；2, tank body；3, guiding device；
31, top convex baffle；31a, filter mesh；32, double horn mouth guide shell above and below middle part；32a, upper double horn mouth；32b, on
Double horn mouth；33, lower porous baffle；34, top snorkel；4, magnetic stirring apparatus；5, cover；6, pillar water leg；6a、
Arc groove；6b, hollow ring cavity；7, carrier probe tube；8, waste liquid probe tube；9, inlet tube；10, outlet tube；11, waste liquid empties
Pipe；12, water inlet pipe；13, outlet pipe；14, cell culture fluid；
In addition, in figure arrow indicate be liquid flow direction.
Specific implementation mode
To make the technical means, the creative features, the aims and the efficiencies achieved by the present invention be easy to understand, with reference to
The drawings and specific embodiments, how the present invention is further explained implements.
A kind of 3D Constituent cells culture carrier provided by the invention, including 3D structure carriers ontology 1, the 3D structure carrier sheets
Body 1 includes at least two-layer nonwoven 1a and at least one layer of holder 1b, and every layer of upper and lower part holder 1b corresponds to and one layer of nothing respectively
Woven fabric 1a is welded and fixed, and every layer of non-woven fabrics 1a can be continuous type nonwoven fabric construct or discontinuous form nonwoven fabric construct；In reality
In, ultrasonic bonding, laser welding, the various welding manners companies such as heat seal welding can be passed through between holder 1b and non-woven fabrics 1a
Connect fixation.
When 1 corresponding every layer of non-woven fabrics 1a of 3D structure carriers ontology is continuous type nonwoven fabric construct, holder 1b therein
Can be arranged to corrugation paper-like or it is latticed in any one, refering to fig. 1 to shown in Fig. 8 and Figure 14.
It is corresponding when 1 corresponding every layer of non-woven fabrics 1a of each 3D structure carriers ontology is discontinuous form nonwoven fabric construct
Every layer of holder 1b be in it is latticed, and every layer of non-woven fabrics 1a therein be made of the non-woven fabric plate of the V-shaped welding of two panels or by
Multiple compositions of the small non-woven fabric plate of setting and V-shaped welding at equal intervals, the small non-woven fabric plate of each V-type welding includes the small nothing of two panels
Woven fabric piece, shown in Fig. 9 to Figure 13.
When holder 1b be corrugation paper-like when, holder 1b use material identical with non-woven fabrics 1a be made and for medical grade without
Woven fabric material, when holder 1b using it is identical with non-woven fabrics 1a when, be more advantageous to cell culture, the nothing between material can be overcome
Malicious compatibility, and cell adherent growth specific surface area can be increased, to improve the stand density of cell；
When holder 1b is latticed, holder 1b is made of medical grade plastic material, for example, polypropylene material.
Every layer of non-woven fabrics 1a thickness therein can be between 0.1mm~100mm any one thickness.
Every layer of holder 1b thickness therein can be between 0.1mm~100mm any one thickness
The cross-sectional shape of 3D structure carriers ontology 1 therein can be any one in circle, ellipse or polygon
Kind；
When the cross-sectional shape of 3D structure carriers ontology 1 is round, the diameter range of 3D structure carriers ontology 1 is 1mm
~10m, altitude range are 1mm~10m；
When the cross sectional shape of 3D structure carriers ontology 1 is ellipse, the major axis dimension of 3D structure carriers ontology 1 is 1mm
~10m, minor axis dimension are 0.9mm~9.9mm, and altitude range is 1mm~10m；
When the cross sectional shape of 3D structure carriers ontology 1 is polygon, the side size range of 3D structure carriers ontology 1 is 1mm
~10m, altitude range are 1mm~10m.
In addition, the cross-sectional area when 3D structure carriers ontology 1 is more than 1962.5mm2When (when 3D structure carriers ontology 1
Cross-sectional shape when being round, the diameter of 3D structure carriers ontology 1 is more than 5cm situations, transversal when 3D structure carriers ontology 1
When face shape is ellipse or polygon, the circumscribed circle diameter of 3D structure carriers ontology 1 is more than 5cm situations) and 3D structure bornes
When corresponding every layer of non-woven fabrics 1a is continuous type nonwoven fabric construct on body ontology 1, in which case, in 3D structure carrier sheets
Several venthole 1c for penetrating through lower surface thereon are further opened on body 1, the diameter range of venthole 1c is 1mm~5cm, ginseng
It reads shown in Figure 15 to Figure 19.
When the cross-sectional area of 3D structure carriers ontology 1 is more than 1962.5mm2When (when the cross of 3D structure carriers ontology 1
When cross sectional shape is round, the diameter of 3D structure carriers ontology 1 is more than 5cm situations, when the cross section shape of 3D structure carriers ontology 1
When shape is ellipse or polygon, the circumscribed circle diameter of 3D structure carriers ontology 1 is more than 5cm situations) and 3D structure carrier sheets
When corresponding every layer of non-woven fabrics 1a is discontinuous form nonwoven fabric construct on body 1, then no longer need to open up on 3D structure carriers ontology 1
Venthole, the self-contained through-hole of net grid support is to be used as cell liquid mass transfer to pass oxygen at this time.
In practical applications, the cross-sectional shape of 3D structure carriers ontology 1 is not limited to disclosed in present specification this
It is several.
Embodiment 1：As depicted in figs. 1 and 2,3D Constituent cells culture carrier provided in this embodiment, by two-layer nonwoven
The 3D structure carriers ontology 1 of 1a and one layer of holder 1b composition, and non-woven fabrics 1a therein is continuous type nonwoven fabric construct, holder 1b
In corrugation paper-like；The cross sectional shape of 3D structure carriers ontology 1 in the present embodiment is that sectional area is less than or equal to 1962.5mm2's
Square.
Embodiment 2：As shown in Figure 3 and Figure 4,3D Constituent cells culture carrier provided in this embodiment is by three layers of nonwoven
The 3D structure carriers ontology 1 of cloth 1a and two layers of holder 1b composition, and every layer of non-woven fabrics 1a therein is continuous type non-woven fabrics knot
Structure, every layer of holder 1b are in corrugation paper-like；With embodiment 1, the cross sectional shape of the 3D structure carriers ontology 1 in the present embodiment is also
Sectional area is less than or equal to 1962.5mm2Square.
Embodiment 3：As shown in Figure 5 and Figure 6,3D Constituent cells culture carrier provided in this embodiment is by two layers of nonwoven
The 3D structure carriers ontology 1 of cloth 1a and one layer of holder 1b composition, and non-woven fabrics 1a therein is continuous type nonwoven fabric construct, holder
1b is in latticed；Same Examples 1 and 2, the cross sectional shape of the 3D structure carriers ontology 1 in the present embodiment be also sectional area be less than or
Equal to 1962.5mm2Square.
Embodiment 4：As shown in Figure 7 and Figure 8,3D Constituent cells culture carrier provided in this embodiment is by three layers of nonwoven
The 3D structure carriers ontology 1 of cloth 1a and two layers of holder 1b composition, and every layer of non-woven fabrics 1a therein is continuous type non-woven fabrics knot
Structure, every layer of holder 1b are in latticed；Same Examples 1 to 3, the cross sectional shape of the 3D structure carriers ontology 1 in the present embodiment
It is less than or equal to 1962.5mm for sectional area2Square.
Embodiment 5：As shown in figure 9,3D Constituent cells culture carrier provided in this embodiment, be by two-layer nonwoven 1a and
The 3D structure carriers ontology 1 of one layer of holder 1b composition, and every layer of non-woven fabrics 1a therein is for discontinuous form nonwoven fabric construct and by two
The non-woven fabric plate of the V-shaped welding of piece forms, and holder 1b is in latticed；Same Examples 1 and 2, the 3D structure carriers in the present embodiment
The cross sectional shape of ontology 1 is also that sectional area is less than or equal to 1962.5mm2Square.
Embodiment 6：As shown in Figure 10 and Figure 11,3D Constituent cells culture carrier provided in this embodiment is by two layers of nothing
The 3D structure carriers ontology 1 of woven fabric 1a and one layer of holder 1b composition, and every layer of non-woven fabrics 1a therein is discontinuous form non-woven fabrics knot
It structure and is made of multiple small non-woven fabric plates that V-shaped welding is arranged at equal intervals, holder 1b is in latticed；Same Examples 1 and 2, this
The cross sectional shape of 3D structure carriers ontology 1 in embodiment is also that sectional area is less than or equal to 1962.5mm2Square
Embodiment 7：As shown in Figure 12 and Figure 13,3D Constituent cells culture carrier provided in this embodiment is by three layers of nothing
The 3D structure carriers ontology 1 of woven fabric 1a and two layers of holder 1b composition, and every layer of non-woven fabrics 1a therein is discontinuous form non-woven fabrics
It structure and is made of multiple small non-woven fabric plates that V-shaped welding is arranged at equal intervals, every layer of holder 1b is in latticed；Same embodiment
1~3, the cross sectional shape of the 3D structure carriers ontology 1 in the present embodiment is also that sectional area is less than or equal to 1962.5mm2Pros
Shape.
Embodiment 8：As shown in figure 14,3D Constituent cells culture carrier provided in this embodiment is by three layers of non-woven fabrics 1a
The 3D structure carriers ontology 1 formed with two layers of holder 1b, and every layer of non-woven fabrics 1a therein is continuous type nonwoven fabric construct,
In mono- layer of holder 1b be in corrugation paper-like, another layer is in latticed；Same Examples 1 to 6, in the present embodiment, 3D structure carrier sheets
The cross sectional shape of body 1 is also that sectional area is less than or equal to 1962.5mm2Square.
Embodiment 9：As shown in figure 15,3D Constituent cells culture carrier provided in this embodiment is by two-layer nonwoven 1a
The 3D structure carriers ontology 1 formed with one layer of holder 1b, and non-woven fabrics 1a therein is continuous type nonwoven fabric construct, holder 1b
In corrugation paper-like；In the present embodiment, the cross sectional shape of 3D structure carriers ontology 1 is that sectional area is more than 1962.5mm2Pros
Shape.
Embodiment 10：As shown in figure 16,3D Constituent cells culture carrier provided in this embodiment is by three layers of non-woven fabrics 1a
The 3D structure carriers ontology 1 formed with two layers of holder 1b, and every layer of non-woven fabrics 1a therein is continuous type nonwoven fabric construct, often
Layer holder 1b is in corrugation paper-like；With embodiment 8, in the present embodiment, the cross sectional shape of 3D structure carriers ontology 1 is also sectional area
More than 1962.5mm2Square.
Embodiment 11：As shown in figure 17,3D Constituent cells culture carrier provided in this embodiment is by two-layer nonwoven 1a
The 3D structure carriers ontology 1 formed with one layer of holder 1b, and non-woven fabrics 1a therein is continuous type nonwoven fabric construct, holder 1b
In latticed；With embodiment 8 and 9, in the present embodiment, the cross sectional shape of 3D structure carriers ontology 1 is also more than for sectional area
1962.5mm2Square.
Embodiment 12：As shown in figure 18,3D Constituent cells culture carrier provided in this embodiment is by three layers of non-woven fabrics 1a
The 3D structure carriers ontology 1 formed with two layers of holder 1b, and every layer of non-woven fabrics 1a therein is continuous type nonwoven fabric construct, often
Layer holder 1b is in latticed；With embodiment 5~7, in the present embodiment, the cross sectional shape of 3D structure carriers ontology 1 is also section
Product is more than 1962.5mm2Square.
Embodiment 13：As shown in figure 19,3D Constituent cells culture carrier provided in this embodiment is by three layers of non-woven fabrics 1a
The D structure carriers ontology 1 formed with two layers of holder 1b, and every layer of non-woven fabrics 1a therein is continuous type nonwoven fabric construct,
In mono- layer of holder 1b be in corrugation paper-like, another layer is in latticed；With embodiment 8~12, in the present embodiment, 3D structure carrier sheets
The cross sectional shape of body 1 is also that sectional area is more than 1962.5mm2Square.
As shown in figure 20, the present invention also provides a kind of bioreactors, and it includes following several components：
Tank body 2；
Several 3D structure carriers ontologies 1 being placed in tank body 2；
The guiding device 3 being vertically installed in tank body 2；
Bottom and the magnetic stirring apparatus 4 positioned at 3 lower part of guiding device are set in tank body 2；
Cover 5 at 2 top of tank body is set；
Pillar water leg 6 in 2 outer bottom of tank body is set；
Wherein, 3D structure carriers ontology 1 is using being above-mentioned to be made of at least two-layer nonwoven 1a and one layer of net grid support 1b；
Wherein, 3 mass activity of guiding device is shelved in tank body 2, is not fixed with tank body 2, can be convenient for picking and placeing in this way, also may be used
Carrier hinge 1 to be filled in tank body 2 provides the effect for passing oxygen, mass transfer, water conservancy diversion；
Wherein, magnetic stirring apparatus 4 is fixed by welding in the inner bottom wall of tank body 2, and by being arranged outside tank body 2
Magnetic force generator drive its operation；The operation of magnetic stirring apparatus 4 is driven to promote the cell training in tank body 2 using magnetic force generator
Nutrient solution 14 flow mode, with tradition stirring motor drive agitating device promote cell culture fluid 14 flow by way of phase
Than this magnetic stirring apparatus 4 eliminates the process for needing to be punched on tank body 2, ensure that the sealing effect of bioreactor tank body
Fruit.Since traditional stirring motor drives agitating device to need to punch on tank body 2, allow the arbor of stirring motor enter tank body with
The arbor that agitating device connected or stretched out agitating device tank body stirring motor connects, and agitating device could be allowed to work, in this way
It can undoubtedly be affected to the leakproofness of tank body in puncture site.
Wherein, cover 5 is tightly connected with 2 top of tank body, in practical applications, thread seal connection or sealing ring can be used
Sealed connection or other connection types, cover 5 and 2 material of tank body can be glass or stainless steel.
Wherein, pillar water leg 6 takes nested mode to connect with 2 outer bottom of tank body, and pillar water leg 6 is mainly used for playing
The effect that support consolidates the effect of tank body 2 and heats or cool down for tank body 2.
Carrier probe tube 7, waste liquid probe tube 8, inlet tube 9, outlet tube 10 and waste liquid evacuated tube have been also extended through on cover 5
11, carrier probe tube 7, waste liquid probe tube 8, inlet tube 9, outlet tube 10 and 11 one end of waste liquid evacuated tube are stretched into inside tank body 2,
The other end stretches out 2 top of tank body；
Wherein, carrier probe tube 7, which stretches into one end of tank body 2, can be deep into 2 lower part of tank body, be mainly used in cell culture
During monitoring, amplification positioned at tank body 2 cell；
Wherein, waste liquid probe tube 8, which stretches into one end of tank body 2, can be deep into the middle part of tank body 2, be mainly used for extracting tank body 2
The waste liquid of interior cell culture, detects remaining nutrition content that it contains and whether cell waste is exceeded；
Wherein, inlet tube 9 stretches into one end of tank body 2 and is generally at the tank mouth of tank body 2, is mainly used for defeated to tank body 2
Enter cell culture fluid 14, the carrier hinge 1 filled in tank body 2 is made to be in suspended state；
Wherein, outlet tube 10 stretches at one end generally also tank mouth in tank body 2 of tank body 2, but slightly below stretches into tank body 2
One end be generally at the tank mouth of tank body 2, be mainly used for export cell culture fluid 14；
Wherein, waste liquid evacuated tube 11, which stretches into one end of tank body 2, would generally be deep into 2 bottom of tank body, be mainly used for tank
Cell waste liquid emptying in body 2.
As shown in figure 21, in the embodiment of this bioreactor, guiding device 3 is by top convex baffle 31, middle part
Lower double horn mouth guide shell 32, lower porous baffle 33 and top snorkel 34 form, wherein double horn mouth is led up and down at middle part
The upper horn mouth 32a of flow cartridge 32 and lower horn mouth 32b are connected to top convex baffle 31 and lower porous baffle 33 respectively, middle part
The lower horn mouth 32b of upper and lower double horn mouth guide shell 32 is also opposite with the paddle of magnetic stirring apparatus 4, top snorkel 34 1
End, which is stretched, to be located inside middle part or more double horn mouth guide shell 32, and the other end is through cover 5 and stretches out 2 top of tank body；It is convex on top
It is further opened with several filter mesh 31a on shape baffle 31, filter mesh 31a is mainly used for retaining carrier hinge 1 and conveniently
Cell culture liquid flows.
Wherein, the convex structure design of top convex baffle 31 has the following advantages that：1) when carrying out cell culture, cell
Culture carrier and cell are not easy to enter lower part from the top of guiding device 3, it is not easy to block filter mesh 31a；2) it can be filled
More cell culture vectors convenient for the amplification of the amplification culture and technique of cell, while also allowing for cell culture vector disengaging tank
Inside body 2, to accurately detect the true growth conditions of 2 inner cell of tank body, and it is convenient for cell sampling and contraposition
The digestion of cell in born of the same parents' culture carrier in tank body 2；3) when carrying out cell culture liquid flowing, the filtering of bigger shuts off face
Product so that cell culture fluid exchanges efficient；
Wherein, it is the advantages of the mouth structure of the double horn up and down design of double horn mouth guide shell 32 above and below middle part：1) loudspeaker on
Mouth 32a can make to form turbulent flow when tank body 2 is flowed in cell culture fluid, make to lead in the cooperation with top convex baffle 31
Entering having with oxygen in tank body 2 higher mixing efficiency and makes mixed effect more preferably；2) water conservancy diversion effect is good, is convenient for each nutritional ingredient
Be sufficiently mixed.
Wherein, the advantages of design of top snorkel 34, is：Increase stroke of the gas in pipeline, increase gas with
The mixability of liquid so that dissolution degree higher of the gas in cell culture fluid body, dissolved oxygen effect are more preferable.
In practical applications, the shape of top convex baffle 31 can be in netted spherical or netted cabochon or netted cone
Shape or other netted convex structures, in the present embodiment, as shown in figure 4, the shape of top convex baffle 31 is then using net
Shape spherical structure.
Wherein, top convex baffle 31, middle part double horn mouth guide shell 32, lower porous baffle 33 and top ventilation up and down
Between pipe 34 it is linked together by welding procedure.
As shown in Figure 20 and Figure 22, to ensure that it is more firm that tank body 2 carries on pillar water leg 6, pressed from both sides in pillar water
6 top of set is additionally provided with the groove 6a to match with 2 exterior bottom wall of tank body, and pillar water leg 6 passes through groove 6a and 2 outer bottom of tank body
Be nested cooperation；
In addition, as shown in figure 22, hollow ring cavity 6b is additionally provided with inside pillar water leg 6, in pillar water leg 3 or so two
It is further respectively had on side wall and the hollow ring cavity 6b water inlet pipes 12 being connected to and outlet pipe 13, wherein water inlet pipe 12 and outlet pipe 13
By being connected to external fluid delivery conduit road indirectly heat or the purpose of cooling can be played for the cell culture fluid 14 in tank body 2.
Its operation principle of bioreactor provided by the invention is as follows：
When work, first cell culture fluid 14 is entered by inlet tube 9 inside tank body 2, is stirred by the magnetic force that bottom is equipped with
Effect and immixture that device 4 generates flow direction (in figure shown in arrow) are mixed, to drive carrier hinge 1 to suspend upwards,
Cell is set to be grown in 1 region of carrier hinge；
During cell culture, the monitoring of carrier probe tube 7 and magnocell culture being equipped with by cover 5 are led to
The waste liquid that waste liquid probe tube 8 extracts cell cultivation process is crossed, nutritional ingredient therein is detected, new convenient for replacement in time or supply
Cell culture fluid is ventilated by top snorkel 34 to the cell culture fluid 14 in tank body 2, to ensure that cell growth is required
Dissolved oxygen etc.；
In the completion of cell culture, the cell culture waste liquid in tank body 2 is excluded by waste liquid evacuated tube 11.
Finally illustrate, the above is only the implementation of the present invention example, is not intended to limit the scope of the invention, it is every
Using equivalent structure or equivalent flow shift made by description of the invention and accompanying drawing content, it is applied directly or indirectly in other
Relevant technical field, is included within the scope of the present invention.
Claims (11)
1. a kind of 3D Constituent cells culture carrier, including 3D structure carriers ontology (1), it is characterised in that：The 3D structure carriers
Ontology (1) includes that at least two-layer nonwoven (1a) and at least one layer of holder (1b), every layer of holder (1b) upper and lower part is divided
Dui Ying not be welded and fixed with one layer of non-woven fabrics (1a), described every layer non-woven fabrics (1a) be continuous type nonwoven fabric construct or
Disconnected type nonwoven fabric construct.
2. 3D Constituent cells culture carrier according to claim 1, it is characterised in that：When each 3D structure carriers sheet
When the corresponding every layer of non-woven fabrics (1a) of body (1) is continuous type nonwoven fabric construct, corresponding every layer of holder (1b) is in corrugation paper-like
Or it is latticed in any one；
It is right when the corresponding every layer of non-woven fabrics (1a) of each 3D structure carriers ontology (1) is discontinuous form nonwoven fabric construct
The every layer of holder (1b) answered is in latticed, and every layer of non-woven fabrics therein (1a) is by the non-woven fabric plate group of the V-shaped welding of two panels
It is formed at or by multiple small non-woven fabric plates of setting and V-shaped welding at equal intervals.
3. 3D Constituent cells culture carrier according to claim 2, it is characterised in that：When the holder (1b) is corrugated paper
When shape, the holder (1b) uses the identical material with the non-woven fabrics (1a) to be made and for medical grade non-woven fabrics material；
When the holder (1b) is latticed, the holder (1b) is made of medical grade plastic material.
4. 3D Constituent cells culture carrier according to claim 1, it is characterised in that：Every layer of non-woven fabrics (1a) thickness
For 0.1mm~100mm；Every layer of holder (1b) thickness is 0.1mm~100mm.
5. 3D Constituent cells culture carrier according to claim 1, it is characterised in that：The 3D structure carriers ontology (1)
Cross-sectional shape be any one in round, ellipse or polygon；
When the cross-sectional shape of the 3D structure carriers ontology (1) is round, the 3D structure carriers ontology (1) it is a diameter of
1mm~10m is highly 1mm~10m；
When the cross sectional shape of the 3D structure carriers ontology (1) is ellipse, the long axis ruler of the 3D structure carriers ontology (1)
Very little is 1mm~10m, and minor axis dimension is 0.9mm~9.9mm, is highly 1mm~10m；
When the cross sectional shape of the 3D structure carriers ontology (1) is polygon, the length of side of the 3D structure carriers ontology (1) is
1mm~10m is highly 1mm~10m.
6. 3D Constituent cells culture carrier according to claim 1, it is characterised in that：When the 3D structure carriers ontology
(1) cross-sectional area is more than 1962.5mm2And corresponding every layer of non-woven fabrics (1a) is continuous on 3D structure carriers ontology (1)
When type nonwoven fabric construct, several ventilations for penetrating through lower surface thereon are further opened on the 3D structure carriers ontology (1)
Hole (1c), and a diameter of 1mm~5cm of the venthole (1c).
7. a kind of bioreactor, it is characterised in that：Include tank body (2) and several such as power being placed in the tank body (2)
Profit requires the 3D structure carriers ontology (1) described in 1；
Also include the guiding device (3) being vertically installed in the tank body (2), guiding device (3) expansion bearing is in institute
It states in tank body (2)；
Also include to be arranged in the interior bottom of the tank body (2) and positioned at the magnetic stirring apparatus of the guiding device (3) lower part
(4), the magnetic stirring apparatus (4) is weldingly fixed in the inner bottom wall of the tank body (2), and by setting tank body (2) outside
The magnetic force generator in portion drives its operation；
Also include the cover (5) being arranged at the top of the tank body (2), the cover (5) connects with sealing at the top of the tank body (2)
It connects；
Also include pillar water leg (6) of the setting in the tank body (2) outer bottom, the pillar water leg (6) and the tank
Body (2) outer bottom nested encryptions.
8. bioreactor according to claim 7, it is characterised in that：Carrier is also extended through on the cover (5) to take
Sample pipe (7), waste liquid probe tube (8), inlet tube (9), outlet tube (10) and waste liquid evacuated tube (11), the carrier probe tube (7),
The tank body (2) inside is stretched into waste liquid probe tube (8), inlet tube (9), outlet tube (10) and waste liquid evacuated tube (11) one end,
The other end stretches out at the top of the tank body (2).
9. bioreactor according to claim 7, it is characterised in that：The guiding device (3) is by top convex baffle
(31), double horn mouth guide shell (32), lower porous baffle (33) and top snorkel (34) form up and down at middle part, in described
Portion up and down double horn mouth guide shell (32) upper horn mouth (32a) and lower horn mouth (32b) respectively with the top convex baffle
(31) and lower porous baffle (33) connection, the middle part up and down double horn mouth guide shell (32) lower horn mouth (32b) also with
The paddle of the magnetic stirring apparatus (4) is opposite, and described top snorkel (34) one end, which is stretched, is located at middle part double horn mouth up and down
Guide shell (32) is internal, and the other end is through the cover (5) and stretches out at the top of tank body (2).
10. bioreactor according to claim 7, it is characterised in that：It is also opened on the top convex baffle (31)
Equipped with several filter mesh (31a), the shape of the top convex baffle (31) is in netted spherical or netted arc surfaced or net
Shape taper, the top convex baffle (31), the upper and lower double horn mouth guide shell (32) in middle part, lower porous baffle (33) and top
Snorkel is linked together between (34) by welding procedure.
11. bioreactor according to claim 7, it is characterised in that：Be equipped at the top of the pillar water leg (6) with
The groove (6a) that tank body (2) exterior bottom wall matches, the pillar water leg (6) pass through the groove (6a) and the tank body
(2) outer bottom nested encryptions；It is additionally provided with hollow ring cavity (6b) inside the pillar water leg (6), in the pillar water leg
(3) water inlet pipe (12) being connected to the hollow ring cavity (3b) and outlet pipe (13) are further respectively had on left and right sides wall.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810905734.2A CN108676716A (en) | 2018-08-10 | 2018-08-10 | A kind of 3D Constituent cells culture carrier and bioreactor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810905734.2A CN108676716A (en) | 2018-08-10 | 2018-08-10 | A kind of 3D Constituent cells culture carrier and bioreactor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108676716A true CN108676716A (en) | 2018-10-19 |
Family
ID=63816421
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810905734.2A Pending CN108676716A (en) | 2018-08-10 | 2018-08-10 | A kind of 3D Constituent cells culture carrier and bioreactor |
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| Country | Link |
|---|---|
| CN (1) | CN108676716A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109609380A (en) * | 2019-02-21 | 2019-04-12 | 江阴新雨生物科技有限公司 | A kind of stereoscopic cell culturing bracket |
| CN109609380B (en) * | 2019-02-21 | 2024-06-04 | 秦皇岛润泽医院有限公司 | Three-dimensional cell culture support |
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| CN1119849A (en) * | 1993-03-25 | 1996-04-03 | 赫汀股份公司 | Fixed-bed bioreactor and carrier bodies for purifying fluids |
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