CN113025491A - Perfusion reactor for in-vitro dynamic culture of cells - Google Patents
Perfusion reactor for in-vitro dynamic culture of cells Download PDFInfo
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- CN113025491A CN113025491A CN202110287690.3A CN202110287690A CN113025491A CN 113025491 A CN113025491 A CN 113025491A CN 202110287690 A CN202110287690 A CN 202110287690A CN 113025491 A CN113025491 A CN 113025491A
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- 230000010412 perfusion Effects 0.000 title claims abstract description 59
- 238000000338 in vitro Methods 0.000 title claims abstract description 14
- 238000010008 shearing Methods 0.000 claims abstract description 28
- 230000002572 peristaltic effect Effects 0.000 claims abstract description 19
- 210000004027 cell Anatomy 0.000 claims abstract description 14
- 239000000017 hydrogel Substances 0.000 claims abstract description 13
- 239000007788 liquid Substances 0.000 claims abstract description 12
- 230000000694 effects Effects 0.000 claims abstract description 10
- 238000004458 analytical method Methods 0.000 claims abstract description 9
- 230000035755 proliferation Effects 0.000 claims abstract description 6
- 239000000463 material Substances 0.000 claims description 9
- 239000012530 fluid Substances 0.000 claims description 7
- 210000003321 cartilage cell Anatomy 0.000 claims description 4
- 230000004663 cell proliferation Effects 0.000 claims description 4
- 238000000034 method Methods 0.000 claims description 4
- 239000011148 porous material Substances 0.000 claims description 3
- 230000002265 prevention Effects 0.000 claims description 3
- 238000012606 in vitro cell culture Methods 0.000 claims description 2
- 210000001612 chondrocyte Anatomy 0.000 abstract description 6
- 238000004113 cell culture Methods 0.000 abstract description 5
- 239000000203 mixture Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 11
- 210000001188 articular cartilage Anatomy 0.000 description 3
- 238000010586 diagram Methods 0.000 description 2
- 229920001296 polysiloxane Polymers 0.000 description 2
- 230000003068 static effect Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000003631 expected effect Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000011343 solid material Substances 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
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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
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
- C12M25/14—Scaffolds; Matrices
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- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/10—Perfusion
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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
- C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
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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
- C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
- C12M35/04—Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
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Abstract
The invention discloses a perfusion reactor for in-vitro dynamic culture of cells, which belongs to the technical field of dynamic cell culture devices and comprises a peristaltic pump, a perfusion chamber and a liquid storage bottle, wherein the peristaltic pump, the perfusion chamber and the liquid storage bottle are connected through a conduit, and a bracket is arranged in the perfusion chamber. According to the perfusion reactor for in-vitro dynamic culture of the cells, the composition structure is simple, the perfusion reactor is easy to build, the hydrogel support is placed in the perfusion chamber in the cell culture process, the culture solution is placed in the solution storage bottle, the peristaltic pump is opened, the flow rate is adjusted, the culture solution is circulated and flows to culture the cells, the shearing force applied to the interior of the support is obtained through finite element analysis, and the shearing force applied to the support can be adjusted by adjusting the flow rate, so that the influence on the proliferation effect of the chondrocytes due to too large or too small shearing force is avoided.
Description
Technical Field
The invention relates to the technical field of dynamic cell culture devices, in particular to a perfusion reactor for in-vitro dynamic cell culture.
Background
The damaged articular cartilage is difficult to self-repair or regenerate, the development of tissue engineering provides possibility for the repair of the articular cartilage, the reason for the extremely poor self-repair capability of the damaged articular cartilage tissue is that no nerve or blood vessel exists in the tissue, different culture modes can cause different morphologies of chondrocytes, and the biological characteristics of the chondrocytes also greatly differ.
The current in vitro culture methods of cells are divided into static culture and dynamic culture. Static culture is generally performed directly from a petri dish. The dynamic culture is divided into three-dimensional hydrogel scaffold culture, various bioreactors and microcarrier culture. The bioreactor comprises a perfusion bioreactor, and the perfusion bioreactor generally forms a closed loop by a perfusion chamber, a peristaltic pump, a liquid storage bottle and a silicone tube. The peristaltic pump provides power, the liquid storage bottle stores culture solution, the bracket is arranged in the perfusion chamber, the culture solution circulates in the whole loop through the power provided by the peristaltic pump, and the flowing culture solution generates fluid shearing force on the chondrocytes. The self-made perfusion bioreactor cannot control the shearing force applied to the interior of the bracket, the hydrogel bracket is broken due to overlarge shearing force, and the cartilage cell proliferation effect of the perfusion bioreactor cannot achieve the expected effect due to the undersize shearing force.
To this end, we propose a perfusion reactor for dynamic in vitro cell culture to solve the above problems.
Disclosure of Invention
The invention aims to solve the problems in the prior art and provides a perfusion reactor for in-vitro dynamic culture of cells.
In order to achieve the purpose, the invention adopts the following technical scheme:
a perfusion reactor for in-vitro dynamic culture of cells comprises a peristaltic pump, a perfusion chamber and a liquid storage bottle, wherein the peristaltic pump, the perfusion chamber and the liquid storage bottle are connected through a conduit, and a support is arranged in the perfusion chamber.
Wherein the peristaltic pump is used for promoting the culture solution to flow in the perfusion reactor.
Wherein the filling chamber is used for placing the hydrogel support.
Wherein, the stock solution bottle is used for holding the culture solution.
Preferably, the conduit is a silicone tube that is gas permeable.
Preferably, the scaffold is a hydrogel scaffold.
Preferably, the scaffold includes a hydrogel scaffold but is not limited to a hydrogel scaffold.
A method for improving cell proliferation effect of a perfusion reactor for dynamic culture comprises the following steps:
step one, calculating the flow velocity inside the perfusion reactor, wherein the flow velocity theoretical calculation formula is as follows:
Vm＝Q/n(D×h)
vm denotes the flow rate, Q denotes the flow rate through the perfusion chamber 2, n denotes the porosity of the hydrogel material, D denotes the width of the perfusion chamber channel, h denotes the thickness of the scaffold;
inputting the Young modulus of the material in the software of the finite element analysis;
inputting the integral shearing force of the bracket calculated in the step one into the software of the finite element analysis, wherein the theoretical calculation formula of the integral shearing force is as follows:
TW＝8υVm/d
TW represents the fluid shear force, upsilon represents the fluid viscosity, Vm represents the flow velocity, and d represents the pore diameter of the stent material;
fourthly, performing true prevention through the finite element analysis software;
fifthly, simulating to obtain the shearing force in the bracket;
and step six, adjusting the flow rate to change the shearing force according to the relation between the shearing force in the bracket obtained in the step five and the flow rate, and improving the proliferation effect of the cartilage cells cultured by the perfusion bioreactor.
Preferably, the finite element analysis software is Ansys software.
The technical scheme provided by the embodiment of the invention has the following beneficial effects:
according to the perfusion reactor for in-vitro dynamic culture of the cells, the composition structure is simple, the perfusion reactor is easy to build, the hydrogel support is placed in the perfusion chamber in the cell culture process, the culture solution is placed in the solution storage bottle, the peristaltic pump is opened, the flow rate is adjusted, the culture solution is circulated and flows to culture the cells, the shearing force applied to the interior of the support is obtained through finite element analysis, and the shearing force applied to the support can be adjusted by adjusting the flow rate, so that the influence on the proliferation effect of the chondrocytes due to too large or too small shearing force is avoided.
Drawings
FIG. 1 is a schematic structural diagram of a perfusion reactor for dynamic in vitro culture of cells according to the present invention;
FIG. 2 is a schematic structural diagram of a perfusion reactor for dynamic in vitro culture of cells according to the present invention.
In the figure: 1. a peristaltic pump; 2. a perfusion chamber; 3. a liquid storage bottle; 4. a conduit.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
Referring to fig. 1-2, a perfusion reactor for in vitro dynamic culture of cells comprises a peristaltic pump 1, a perfusion chamber 2 and a liquid storage bottle 3, wherein the peristaltic pump 1, the perfusion chamber 2 and the liquid storage bottle 3 are connected through a conduit 4, and a bracket is arranged in the perfusion chamber 2.
More specifically, the scaffold is a hydrogel scaffold.
A method for improving cell proliferation effect of a perfusion reactor for dynamic culture comprises the following steps:
step one, calculating the flow velocity inside the perfusion reactor, wherein the theoretical calculation formula of the flow velocity is as follows:
Vm＝Q/n(D×h)
vm denotes the flow rate, Q denotes the flow rate through the perfusion chamber 2, n denotes the porosity of the hydrogel material, D denotes the width of the perfusion chamber channel, h denotes the thickness of the scaffold;
inputting the Young modulus of the material in software of finite element analysis;
step three, inputting the integral shearing force of the bracket calculated in the step one in software of finite element analysis, wherein a theoretical calculation formula of the integral shearing force is as follows:
TW＝8υVm/d
TW represents the fluid shear force, upsilon represents the fluid viscosity, Vm represents the flow velocity, and d represents the pore diameter of the stent material;
step four, performing true prevention through finite element analysis software;
fifthly, simulating to obtain the shearing force in the bracket;
and step six, adjusting the flow rate to change the shearing force according to the relation between the shearing force in the bracket obtained in the step five and the flow rate, and improving the proliferation effect of the cartilage cells cultured by the perfusion bioreactor.
Wherein the flow rate inside the perfusion reactor is increased by a peristaltic pump when the resulting shear force inside the stent is below a threshold range.
Wherein the flow rate inside the perfusion reactor is reduced by a peristaltic pump when the resulting shear force inside the stent exceeds a threshold range.
Where young's modulus is a physical quantity describing the ability of a solid material to resist deformation, also called tensile modulus.
More specifically, the finite element analysis software is Ansys software.
The perfusion reactor is a self-made perfusion reactor and mainly comprises a peristaltic pump 1, a perfusion chamber 2, a liquid storage bottle 3 and a conduit 4, power is provided by the peristaltic pump 1, so that a culture solution can circulate in a closed loop, the shearing force applied to the inside of a support can be obtained through finite element analysis, the shearing force applied to the support can be adjusted by adjusting the flow rate, and the influence on the proliferation effect of chondrocytes due to too large or too small shearing force is avoided.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (4)
1. A perfusion reactor for in-vitro dynamic culture of cells is characterized by comprising a peristaltic pump (1), a perfusion chamber (2) and a liquid storage bottle (3), wherein the peristaltic pump (1), the perfusion chamber (2) and the liquid storage bottle (3) are connected through a conduit (4), and a support is arranged in the perfusion chamber (2).
2. A perfusion reactor for dynamic in vitro cell culture according to claim 1, wherein the scaffold is a hydrogel scaffold.
3. A method for improving cell proliferation effect of perfusion reactor for dynamic culture according to any of claims 1 or 2, comprising the following steps:
step one, calculating the flow velocity inside the perfusion reactor, wherein the flow velocity theoretical calculation formula is as follows:
Vm＝Q/n(D×h)
vm denotes the flow rate, Q denotes the flow rate through the perfusion chamber (2), n denotes the porosity of the hydrogel material, D denotes the width of the perfusion chamber channel, h denotes the thickness of the scaffold;
inputting the Young modulus of the material in the software of the finite element analysis;
inputting the integral shearing force of the bracket calculated in the step one into the software of the finite element analysis, wherein the theoretical calculation formula of the integral shearing force is as follows:
TW＝8υVm/d
TW represents the fluid shear force, upsilon represents the fluid viscosity, Vm represents the flow velocity, and d represents the pore diameter of the stent material;
fourthly, performing true prevention through the finite element analysis software;
fifthly, simulating to obtain the shearing force in the bracket;
and step six, adjusting the flow rate to change the shearing force according to the relation between the shearing force in the bracket obtained in the step five and the flow rate, and improving the proliferation effect of the cartilage cells cultured by the perfusion bioreactor.
4. The method for calculating the shear force of a perfusion reactor for dynamic in vitro culture of cells according to claim 3, wherein the finite element analysis software is Ansys software.
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| CN202110287690.3A CN113025491A (en) | 2021-03-17 | 2021-03-17 | Perfusion reactor for in-vitro dynamic culture of cells |
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2005287162A1 (en) * | 2004-09-16 | 2006-03-30 | Becton, Dickinson And Company | Perfusion bioreactor for culturing cells |
| CN102154101A (en) * | 2011-01-11 | 2011-08-17 | 王林 | Integrated dynamic inoculation culture method for osteogenous seed cell and matched bioreactor thereof |
| CN103290145A (en) * | 2013-06-03 | 2013-09-11 | 中山大学 | Pouring type bioreactor experiment method and device used by same |
| CN205398649U (en) * | 2016-03-08 | 2016-07-27 | 兰青 | 3D prints perfusion culture apparatus who contains cell aquogel support |
| CN106126869A (en) * | 2016-08-23 | 2016-11-16 | 中国石油大学(华东) | The numerical value simplified calculation method of ground liquid in storage tank coupled mode |
| WO2017152343A1 (en) * | 2016-03-07 | 2017-09-14 | 浙江大学 | Recirculating perfusion bioreactor device that can realize three-dimensional scaffold recirculating perfusion |
| CN108728360A (en) * | 2018-06-09 | 2018-11-02 | 上海天引生物科技有限公司 | A kind of devices and methods therefor that cell co-cultures |
| CN111876329A (en) * | 2020-08-31 | 2020-11-03 | 大连理工大学 | Immune isolation dynamic co-culture bioreactor for hematopoietic stem cell in-vitro culture |
-
2021
- 2021-03-17 CN CN202110287690.3A patent/CN113025491A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2005287162A1 (en) * | 2004-09-16 | 2006-03-30 | Becton, Dickinson And Company | Perfusion bioreactor for culturing cells |
| CN102154101A (en) * | 2011-01-11 | 2011-08-17 | 王林 | Integrated dynamic inoculation culture method for osteogenous seed cell and matched bioreactor thereof |
| CN103290145A (en) * | 2013-06-03 | 2013-09-11 | 中山大学 | Pouring type bioreactor experiment method and device used by same |
| WO2017152343A1 (en) * | 2016-03-07 | 2017-09-14 | 浙江大学 | Recirculating perfusion bioreactor device that can realize three-dimensional scaffold recirculating perfusion |
| CN205398649U (en) * | 2016-03-08 | 2016-07-27 | 兰青 | 3D prints perfusion culture apparatus who contains cell aquogel support |
| CN106126869A (en) * | 2016-08-23 | 2016-11-16 | 中国石油大学(华东) | The numerical value simplified calculation method of ground liquid in storage tank coupled mode |
| CN108728360A (en) * | 2018-06-09 | 2018-11-02 | 上海天引生物科技有限公司 | A kind of devices and methods therefor that cell co-cultures |
| CN111876329A (en) * | 2020-08-31 | 2020-11-03 | 大连理工大学 | Immune isolation dynamic co-culture bioreactor for hematopoietic stem cell in-vitro culture |
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Application publication date: 20210625 |