CN106754627A - A kind of carrier for cultured cells - Google Patents
A kind of carrier for cultured cells Download PDFInfo
- Publication number
- CN106754627A CN106754627A CN201710107175.6A CN201710107175A CN106754627A CN 106754627 A CN106754627 A CN 106754627A CN 201710107175 A CN201710107175 A CN 201710107175A CN 106754627 A CN106754627 A CN 106754627A
- Authority
- CN
- China
- Prior art keywords
- carrier
- cultured cells
- cell
- cells according
- core
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 210000004748 cultured cell Anatomy 0.000 title claims abstract description 21
- 239000000463 material Substances 0.000 claims abstract description 32
- 238000004113 cell culture Methods 0.000 claims abstract description 22
- 239000007788 liquid Substances 0.000 claims abstract description 15
- 239000000203 mixture Substances 0.000 claims abstract description 7
- 238000012545 processing Methods 0.000 claims abstract description 5
- 238000007493 shaping process Methods 0.000 claims abstract description 5
- 239000002991 molded plastic Substances 0.000 claims abstract description 3
- 238000012946 outsourcing Methods 0.000 claims abstract description 3
- 230000005291 magnetic effect Effects 0.000 claims description 29
- 238000007789 sealing Methods 0.000 claims description 14
- 239000007769 metal material Substances 0.000 claims description 8
- 239000002245 particle Substances 0.000 claims description 7
- 239000012530 fluid Substances 0.000 claims description 6
- 230000002209 hydrophobic effect Effects 0.000 claims description 3
- 239000002184 metal Substances 0.000 claims description 3
- 229910052751 metal Inorganic materials 0.000 claims description 3
- 239000004033 plastic Substances 0.000 claims description 3
- 229920003023 plastic Polymers 0.000 claims description 3
- 238000003466 welding Methods 0.000 claims description 3
- 238000005275 alloying Methods 0.000 claims description 2
- 239000000470 constituent Substances 0.000 claims description 2
- 239000003292 glue Substances 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 238000001746 injection moulding Methods 0.000 claims description 2
- 229920000344 molecularly imprinted polymer Polymers 0.000 claims description 2
- 230000005298 paramagnetic effect Effects 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 239000000969 carrier Substances 0.000 claims 2
- 210000004027 cell Anatomy 0.000 abstract description 48
- 230000012010 growth Effects 0.000 abstract description 13
- 238000004519 manufacturing process Methods 0.000 abstract description 8
- 238000003756 stirring Methods 0.000 abstract description 8
- 230000006872 improvement Effects 0.000 abstract description 2
- -1 polypropylene Polymers 0.000 description 10
- 239000002609 medium Substances 0.000 description 9
- 238000010586 diagram Methods 0.000 description 8
- SZVJSHCCFOBDDC-UHFFFAOYSA-N ferrosoferric oxide Chemical compound O=[Fe]O[Fe]O[Fe]=O SZVJSHCCFOBDDC-UHFFFAOYSA-N 0.000 description 8
- 239000004810 polytetrafluoroethylene Substances 0.000 description 8
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 8
- 238000000034 method Methods 0.000 description 7
- 239000004425 Makrolon Substances 0.000 description 5
- 239000004743 Polypropylene Substances 0.000 description 5
- 239000004793 Polystyrene Substances 0.000 description 5
- 230000010261 cell growth Effects 0.000 description 5
- 239000001963 growth medium Substances 0.000 description 5
- 235000015097 nutrients Nutrition 0.000 description 5
- 229920000515 polycarbonate Polymers 0.000 description 5
- 229920001155 polypropylene Polymers 0.000 description 5
- 229920002223 polystyrene Polymers 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 230000008901 benefit Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 229940056319 ferrosoferric oxide Drugs 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 229910000976 Electrical steel Inorganic materials 0.000 description 3
- 230000001464 adherent effect Effects 0.000 description 3
- 239000000919 ceramic Substances 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 238000004381 surface treatment Methods 0.000 description 3
- 108010010803 Gelatin Proteins 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000013019 agitation Methods 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000007667 floating Methods 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 238000005342 ion exchange Methods 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 229920002635 polyurethane Polymers 0.000 description 2
- 239000004814 polyurethane Substances 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 238000004114 suspension culture Methods 0.000 description 2
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 101100230376 Acetivibrio thermocellus (strain ATCC 27405 / DSM 1237 / JCM 9322 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372) celI gene Proteins 0.000 description 1
- 229910000967 As alloy Inorganic materials 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- GKWTZACONBQTNK-IVIZKJKWSA-N N-benzyl-7H-purin-6-amine (1R,2R,5R,8R,9S,10R,12S)-12-hydroxy-11-methyl-6-methylidene-16-oxo-15-oxapentacyclo[9.3.2.15,8.01,10.02,8]heptadecane-9-carboxylic acid (1R,2R,5R,8R,9S,10R,12S)-12-hydroxy-11-methyl-6-methylidene-16-oxo-15-oxapentacyclo[9.3.2.15,8.01,10.02,8]heptadec-13-ene-9-carboxylic acid Chemical compound C(Nc1ncnc2nc[nH]c12)c1ccccc1.CC12[C@H]3[C@H](C(O)=O)[C@@]45C[C@@H](CC[C@H]4[C@@]3(CC[C@@H]1O)OC2=O)C(=C)C5.CC12[C@H]3[C@H](C(O)=O)[C@@]45C[C@@H](CC[C@H]4[C@]3(OC1=O)C=C[C@@H]2O)C(=C)C5 GKWTZACONBQTNK-IVIZKJKWSA-N 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 108010039918 Polylysine Proteins 0.000 description 1
- 229920005830 Polyurethane Foam Polymers 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- FJWGYAHXMCUOOM-QHOUIDNNSA-N [(2s,3r,4s,5r,6r)-2-[(2r,3r,4s,5r,6s)-4,5-dinitrooxy-2-(nitrooxymethyl)-6-[(2r,3r,4s,5r,6s)-4,5,6-trinitrooxy-2-(nitrooxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-3,5-dinitrooxy-6-(nitrooxymethyl)oxan-4-yl] nitrate Chemical compound O([C@@H]1O[C@@H]([C@H]([C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O)O[C@H]1[C@@H]([C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@@H](CO[N+]([O-])=O)O1)O[N+]([O-])=O)CO[N+](=O)[O-])[C@@H]1[C@@H](CO[N+]([O-])=O)O[C@@H](O[N+]([O-])=O)[C@H](O[N+]([O-])=O)[C@H]1O[N+]([O-])=O FJWGYAHXMCUOOM-QHOUIDNNSA-N 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 239000010868 animal carcass Substances 0.000 description 1
- 230000003466 anti-cipated effect Effects 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000012832 cell culture technique Methods 0.000 description 1
- 230000005779 cell damage Effects 0.000 description 1
- 208000037887 cell injury Diseases 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 229910017052 cobalt Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 235000012489 doughnuts Nutrition 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- XWHPIFXRKKHEKR-UHFFFAOYSA-N iron silicon Chemical compound [Si].[Fe] XWHPIFXRKKHEKR-UHFFFAOYSA-N 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 238000012007 large scale cell culture Methods 0.000 description 1
- 238000003754 machining Methods 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 229910001004 magnetic alloy Inorganic materials 0.000 description 1
- 238000010907 mechanical stirring Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 229910052759 nickel Inorganic materials 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 229910000889 permalloy Inorganic materials 0.000 description 1
- 229920000656 polylysine Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 239000011496 polyurethane foam Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 238000005476 soldering Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 210000003501 vero cell Anatomy 0.000 description 1
- 229960004854 viral vaccine Drugs 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/0068—General culture methods using substrates
- C12N5/0075—General culture methods using substrates using microcarriers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2531/00—Microcarriers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
Abstract
The invention belongs to field of cell culture, specifically related to a kind of carrier for cultured cells, the carrier is to be shaped to real core or hollow body by homogenous material or mixture material integral molded plastic, or real core or hollow body are shaped to by homogenous material or mixture material overall processing, or real core or hollow body are made up of outsourcing watchcase, built-in core thing etc., or real core or hollow body are constituted by carrier element, built-in core thing, capping etc..A closed entirety is formed after whole carrier shaping, mainly carrier outer wall is supplied to cell apposition growth or realizes the function of liquid in carrier stirring cell culture container.The present invention greatly improves the manufacturing cost of the carrier of cultured cells by the improvement in structure, optimizes the difficulty of cell culture, conveniently can quickly and easily realize the propagation of polytype cell, it is possible to scale, large batch of cultured cells.
Description
Technical field
The present invention relates to field of cell culture, a kind of carrier for cell culture.
Background technology
Cell culture refers to zooblast isolated growth and propagation under proper culture conditions, and keep its feature and
The technology of function.Coming from the most cells of animal carcasses tissue needs adherent monolayers to grow.As long as they are not converted to
Non-adherent dependence, it is necessary to be attached on suitable solid dielectric and sprawl, growth could be started.
Traditional cell culture is cultivated using Tissue Culture Dish, and culture dish is suitable to cell attachment and life by treatment
Long, a traditional culture dish provides 78.52cm2And 10 can be supported6-107Individual cell attachment growth.Then, Ren Menkai
Beginning uses the cell spinner bottle, the device such as roller bottle and doughnut to carry out cell culture.This kind of method technique labour intensity is big, carefully
Born of the same parents yield poorly, and product differences between batches are big, and easily cause pollution, take time and effort, and can not meet cell is produced in society now
The demand of thing high-volume culture.
Microcarrier is that a class used in cell culture is non-toxic, non-rigid, density is homogeneous, transparent little particle.Energy
Anchorage-dependent cell is attached to particle surface carries out suspension culture, so as to increase the area of cell attachment growth, is conducive to
The large-scale culture of cell and collection.
The use of Microcarrier Culture Techniques is earliest Van Wezel in 1967 using ion-exchange gel (DEAE-
Sephadex A50) used as carrier, mild agitation can make the ion-exchange gel be suspended in culture medium as small particle
In, because this microcarrier carries electric charge, beneficial to cell attachment in carrying out growth and breeding on carrier, and carrier is in suspension
State can not only greatly increase cell bond area, cell is fully contacted with culture medium again, and then be conducive to the growth of cell,
Therefore the purpose of mass propgation cell can be reached.
Microcarrier culture is most rising a kind of zooblast large-scale culture technology generally acknowledged at present, due to it
Have the advantage of suspension culture and adhere-wall culture concurrently, and amplify easy, be widely used in and cultivate all kinds cell, production vaccine,
Protein product, such as 293 cells, sarcoblast, Vero cells, Chinese hamster ovary celI.Good cell culture microcarrier has following
Feature：Surface area/volume (S/V) is big, and the cell yield of unit volume nutrient solution is high；Suspending, culture and adhere-wall culture fusion exist
Together, both advantages are had concurrently；Can with simple microscope observation of cell bead surface growing state；Simplify cell life
The detection and control of the various environmental factors of length, favorable reproducibility；Culture medium utilization rate is higher；Easily it is amplified culture；Cell is received
Obtain process uncomplicated；Labour intensity is small；Culture systems floor space and space are small.Various commercialized micro- loads are had been developed at present
Body, including dextran microcarrier, polylysine liquid microcarrier, Gelatin based Macroporous Microcarriers, cellulose microcarriers, the micro- load of shitosan
Body, chitin microcarrier, polystyrene microcarrier, polyurethane foam microcarrier, alginate jelly microcarrier and the micro- load of magnetic
Body etc..
Large-scale cell culture is effective way that viral vaccine or protein product are produced by mammaliancellculture
Footpath.Anchorage-dependent cell is attached on microcarrier and suspends culture as by far the most commonly used, maximally effective using bioreactor
One of cell large-scale culture pattern.
Commercialization microcarrier currently used in the market mainly has：As the Cytodex I of GE companies, Cytodex II,
Cytodex III and the Cytopore of Thermo companies, Cytoline, Biosilon, CultispherG etc., the microcarrier of the above
Because its making needs mutually reciprocal by various biological respinse solvents, macromolecule polymer material and additive, reducing agent etc.
, should neutralize and clean, operation is various, complex process, so expensive, current market price has reached ten thousand yuan of per kilogram 3-10
, also there is supply of material phenomenon not in time often in RMB.And because the size of such microcarrier is at several microns to hundreds of microns
, volume is small, needs substantial amounts of microcarrier to insert in reactor in cell large-scale culture, easily causes microcarrier heap
Product so that the space very little between connected microcarrier, cell density than larger, bottom and centre cell just
It is difficult to the adequate nutrients needed for obtaining cell propagation, and the waste discharge that influence cell consumption is crossed so that the growth ring of cell
Border and growth conditions are deteriorated, and the cell growth for easily causing bottom is slow or even dead.Dead cell is damaged to be taken off from carrier
Drop into and influence the growth of other cells.If stem cell, then non-specific differentiation is more prone to.
Therefore, the carrier for being suitable in field of cell culture used by many animals cell large-scale culture is developed, is had
Prospect is widely applied, the development to promoting bioreactor carrier mass cell culture technique is anticipated with important history
Justice.
The purpose of the present invention is, for the Multiplying culture of biological cell provides a kind of growing carrier, to possess it bigger
Cell Absorption Growth space, and space between carrier forms the microenvironment of suitable cell growth, can be with large-scale culture institute
The cell for needing.And the function of stir culture base is realized with the motion of culture vessel or reactor, it is to avoid due to traditional anti-
Device is answered to produce shearing force using mechanical stirring theory, it is so as to cause cell damage or even dead.
In addition it is also possible to increase magnetisable material to carrier, make it possible to control the motion of carrier by external magnetic field, divide
Cloth and state, realize stir culture base fluid body evenly and the more excellent cell growth microenvironment of formation.
The present invention has the advantages that structure and manufacturing process are simple and convenient to operate, it is adaptable to biological as large-scale culture
The growing carrier of cell.It is characterized in：During carrier is inserted into culture vessel or reactor, by magnetic field or by its buoyancy
Or gravity so that carrier floats in culture vessel or reactor, sinks or suspends, and agitation culture in the process
Based sols, make nutrient full and uniformization in liquid, can provide good Metabolic support for the suspension cell of culture, it is possible to
Allow attached cell to be easier absorption to be grown on carrier, bred.
The content of the invention
The invention discloses a kind of new carrier for cell culture.
The carrier of a kind of new cell culture involved in the present invention, it is mainly by homogenous material or mixture material
Integral molded plastic is shaped to real core or hollow body, or is shaped to real core or hollow by homogenous material or mixture material overall processing
Body, or real core or hollow body are made up of outsourcing watchcase, built-in core thing etc., or by structures such as carrier element, built-in core thing, cappings
Cheng Shixin or hollow body.Characterized in that, forming a closed entirety, the size value scope of carrier after whole carrier shaping
For：More than or equal to 1mm, less than the inwall length of side of culture vessel；This feature is allowed to differ markedly from the micro- load in other inventions
Body, it is mainly carrier outer wall and is supplied to cell apposition growth, realizes that carrier stirs the function of liquid in cell culture container simultaneously
It is uniformly distributed suspension cell.
Above-described carrier can be combined by multiple constitutional details and formed a whole body, each part using secondary or
Multi-step injection molding connects into the entirety of sealing, or is bonded into an entirety for sealing using glue, or is connected into one using supersonic welding
The entirety of individual sealing, or an entirety for sealing is pressed into using interference fit in structure, or screw up into one using screw thread
The entirety of sealing, or use laser sintered into the entirety for sealing, or entirety for sealing etc. is heated into respectively using heating
Plant combination.
Above-described carrier is to be shaped as spherical or elliposoidal or patty or possess the smooth without rib of polygonal profile
Angle object.
The optimal size value scope of above-described carrier is：More than or equal to 1mm, but less than the inwall side of culture vessel
It is long.
The outer wall surface of above-described carrier can be fabricated to shiny surface, or can be fabricated to round and smooth concave-convex curved surface, or
Various curved forms such as corrugated surface can be fabricated to.
Density after above-described carrier shaping should meet with the density ratio of medium liquid：0<d<2, d is carrier
Density and medium liquid density ratio, optimal ratio range is：0.95<d<1.05, to ensure it in culture vessel
Can realize floating, sink or being suspended in medium liquid with natural buoyancy or weight in nutrient solution.
Above-described built-in core thing can by permalloy, silicon-iron soft magnetic alloy, ferroso-ferric oxide and iron, cobalt, nickel and its
The magnetic conductivity such as alloy paramagnetic metal material high is made.To ensure that carrier in the environment of no magnetic field, does not have magnetic, so that
The aggregation capability that attracts each other will not be produced.And in the environment of having magnetic field, can be moved by Magnetic Control
Above-described built-in core thing can be embedded at carrier element end face opening, or can be placed on recessed in carrier element
Trench bottom, carrier element top seal is inlaid in by capping.
Above-described built-in core thing is made up of single bulk, sheet or particle, or by multiple block, sheets or particle
Composition, or by powder constituent, can be moved according to magnetic direction.
Above-described carrier element material can use polystyrene, polypropylene, makrolon, PBS resins, polytetrafluoro
The plastic materials such as ethene, Kynoar, cellulose nitrate, polyurethane, it would however also be possible to employ the non-metallic material such as glass, ceramics, also
Can be using metal or alloying metal materials such as stainless steels.
When simultaneously cultured cells in cell culture container is inserted, the size value of carrier should meet x to above-described carrier<L/
2, wherein x are any path length (such as the diameter of circular pearl, the most major diameter or most minor axis of ellipse pearl) of carrier, and L is culture
Any one length in two most short length of sides in the die cavity of container inside, when the size of the length of side can meet loading multiple carrier,
Its size value scope should meet below equation：L/n>x>L/(n+1/2)；Optimal size value scope is；L/(n+1/10)>x>L/
(n+1/3).N is the number of carrier.
Above-described carrier can use hydrophilic or hydrophobic treatment, or surface attachment Thermo-sensitive Molecularly Imprinted Polymer on surface
Deng material.
The present invention relative to the existing carrier of in the market, with following some advantages：
1., simple structure of the present invention, convenient processing and manufacture, production technology is easy, low cost, it is possible to achieve scale, big
The manufacturing of batch.
2. it is, of the invention because processing and manufacturing cost are low, using single use mode cultured cells, without repeating
It is used for multiple times, so as to avoid producing the problem of residual contamination, cleaning inconvenience and sterilizing.
3., the present invention can be in cell culture container or reactor, by magnetic field or weight and culture medium solution
Proportion realizes the function of stirring with this come the function of realizing floating, sink and suspending in culture medium solution.
4. it is, of the invention because its body dimension is than larger, in more than 1mm, can be in mass cell culture, carrier heap
After product, larger space can be formed between carrier, meet the growth of suspension cell and the exchange of nutrient needed for cells grown.
The present invention greatly improves the manufacturing cost of the carrier of cultured cells by the improvement in structure, optimizes
The difficulty of cell culture, conveniently can quickly and easily realize the propagation of polytype cell, it is possible to scale, large batch of culture
Cell.
Brief description of the drawings
Fig. 1 is the cross section structure schematic diagram of single material ball type carrier of the invention.
Fig. 2 is the cross section structure schematic diagram of hollow ball shape magnetic conduction carrier of the present invention.
Fig. 3 is the cross section structure schematic diagram of compounded magnetic conductive ball type carrier of the present invention.
Fig. 4 is the cross section structure schematic diagram of present invention surface treatment ball type carrier.
Specific embodiment
Embodiments of the invention hereinafter will be described in detail with reference to the accompanying drawing of the above, with reference to these accompanying drawings, wherein whole
In individual view, similar reference characteristic is appointed as similar or corresponding part.In embodiment in the present invention, which judges
For purport of the invention can be made to cause unnecessary fuzzy known ability and the detailed description of structure to be ignored.
The embodiment of the above-described carrier for cell culture, is, in order to be further illustrated to of the invention, should not
It is interpreted as being completely covered the content included by the present invention.And the species of the material for limiting all structure of container parts is not used in, with
And the limitation of the institute's basic parameter defined in for the present embodiment.
Embodiment one：
The cross section structure schematic diagram of single material ball type carrier as shown in Figure 1, the ball type carrier of single material can be wrapped
Include：Solid ball type carrier 1 and hollow ball shape carrier 2.Solid ball type carrier 1 it is characterized in that：Using polystyrene or polypropylene
Or the injection of the polymeric material such as makrolon or polytetrafluoroethylene (PTFE) or machining turn into solid sphere, can also use or glass,
The non-metallic materials such as ceramics are processed into solid sphere.A diameter of 6mm of spheroid, due to its overall density ratio medium liquid
Density it is big, so in reactor or culture vessel, the carrier can be deposited in the bottom of medium liquid.Hollow ball shape carrier
2 it is characterized in that：Using polystyrene or the injection of the polymeric material such as polypropylene or makrolon or polytetrafluoroethylene (PTFE) or machinery
Hollow ball is processed into, can also be used or the non-metallic material such as glass, ceramics is processed into hollow ball.Spheroid it is a diameter of
6mm, because the density of its overall density ratio medium liquid is small, carrier is cultivated in being suspended in reactor or culture vessel
Above base fluid body., it is necessary to reactor or culture vessel can be realized rotating or swinging in incubation, carrier can be made upper and lower
Left and right or by setting track move.
Embodiment two：
The cross section structure schematic diagram of hollow ball shape magnetic conduction carrier as shown in Figure 2, it is characterised in that：Mainly by shell 1
Constituted with inner core 2.Shell 1 is using polystyrene or the polymeric material such as polypropylene or makrolon or polytetrafluoroethylene (PTFE) as in
Between for hollow sphere shell, inner core 2 using with magnetic property ferroso-ferric oxide or silicon steel sheet.Inner core 2 is inserted outside
In space inside shell 1.A diameter of 6mm, a diameter of 4mm of internal diameter hollow of whole spheroid.Carrier is by inserting ferroso-ferric oxide
Or the weight of silicon steel sheet controls the weight of whole carrier, make what its overall density was contrasted relative to liquid medium volume density
Size controls carrier to be floated in medium liquid, suspended or sinks.Make whole carrier in the environment of having magnetic field, can
Adherent cell is realized, and carrier can adsorb the magnetic pole in reactor or culture vessel.Carrier is transported in medium liquid
When dynamic, stir culture base fluid body can be played a part of, while the fertile surface area of attached cell can be increased, improve culture
Efficiency.
Embodiment three：
The cross section structure schematic diagram of compounded magnetic conductive ball type carrier as shown in Figure 3, it is characterised in that：It is main by spherical cap 1,
Capping 2, magnetic core 3 and spheroid 4 are constituted.In the space that magnetic core 3 is inserted in the middle of spheroid 4, then capping 2 is covered on the upper surface of spheroid 4,
The space in spheroid 4 is set to form closed space, then that spherical cap pushed down into capping is 2-in-1 on spheroid 4, carrier is formed a circle
Closed entirety.Spherical cap 1 and spheroid 4 can be by makrolon, polypropylene, PBS resins, polytetrafluoroethylene (PTFE), Kynoar, nitric acid
The material injections such as fiber, polyurethane are molded.Magnetic core 3 is made of magnetically permeable ferroso-ferric oxide or silicon steel sheet.Capping 2 is in not shadow
Can be processed into using plastic material injection or the material such as metal, nonmetallic in the case of ringing weight.Can also use and magnetic
The identical material of core 3, increases magnetic.Spherical cap 1 can be moulded on spheroid 4 using overall parcel capping 2；Can also be using ultrasound
Wave soldering is connected on spheroid 4；Spherical cap 1 and spheroid 4 can also be made to bond together using laser sintered, form a closed load
Body；Can also be heated using overall, spherical cap 1 and the welding of spheroid 4 is formed closed entirety.Carrier can be with magnetic field environment
The change in magnetic field and move, so as to realize the function of stir culture base fluid body in reactor or culture vessel, and can conduct
The attachment of adherent cell growth, propagation so that cell obtains maximized culture effect under same environment, improves efficiency.
Example IV：
The cross section structure schematic diagram of surface treatment ball type carrier as shown in Figure 4, it is characterised in that：Mainly by hollow ball
Body 1 and face coat 2 are constituted.Hollow ball 1 is made up of the material for being mixed with magnetic conductivity, then in the surface attachment one of hollow ball 1
The avirulent process layer of layer.Surface treatment can use plasma hydrophilic or hydrophobic treatment, it would however also be possible to employ attachment Thermo-sensitive point
Sub- polymer, it is also possible to surface attachment gelatin or collagen etc..Treated carrier surface is non-toxic, is adapted to attached cell life
Long, propagation.Simultaneously because the hollow ball 1 of nexine carries magnetic conduction property, can be in the magnetic field in reactor or culture vessel
Motion, while stir culture base fluid body, as the growth attachment of attached cell, increases the surface area of cell culture, carries
Culture efficiency high.
The embodiment of above-described carrier, is that, for the further illustrating in cell culture to the present invention, should not manage
Solve as the content included by the present invention is completely covered.And the species of the material for limiting all carrier structure parts is not used in, and
The limitation of the institute's basic parameter defined in for the present embodiment.
Claims (12)
1. a kind of carrier for cultured cells, is to be shaped to real core or hollow by homogenous material or mixture material integral molded plastic
Body, or real core or hollow body are shaped to by homogenous material or mixture material overall processing, or by outsourcing watchcase, built-in
Core thing etc. constitutes real core or hollow body, or constitutes real core or hollow body by carrier element, built-in core thing, capping；Its feature
It is to form a closed entirety after whole carrier shaping, the size value scope of described carrier is：More than or equal to 1mm；Carry
The density of body should meet with the density ratio of medium liquid：0<d<2, d is support density and medium liquid density ratio.
2. the carrier for cultured cells according to claim 1, it is characterised in that combined by multiple constitutional details and constituted
One overall body, each part connects into the entirety of sealing using secondary or Multi-step injection molding, or is bonded into one using glue
The entirety of sealing, or an entirety for sealing is connected into using supersonic welding, or using interference fit it is pressed into one in structure
The entirety of sealing, or an entirety for sealing is screwed up into using screw thread, or using laser sintered into an entirety for sealing, or adopt
The various combinations such as an entirety for sealing are heated into heating.
3. the carrier for cultured cells according to claim 1, it is characterised in that support shapes are spherical or elliposoidal
Or patty or possess the smooth without corner angle object of polygonal profile.
4. the carrier for cultured cells according to claim 1, it is characterised in that the size value scope of carrier be more than
Equal to 1mm, less than or equal to 10mm.
5. the carrier for cultured cells according to claim 1, it is characterised in that the outer wall surface of carrier is smooth
Face, or round and smooth concave-convex curved surface, or corrugated surface form.
6. the carrier for cultured cells according to claim 1, it is characterised in that density and culture after carrier shaping
The density ratio of base fluid body is：0.95<d<1.05.
7. the carrier for cultured cells according to claim 1, it is characterised in that built-in core thing magnetic conductivity paramagnetic high
Property metal material is made.
8. the carrier for cultured cells according to claim 1, it is characterised in that built-in core thing is embedded in carrier element end
At the opening of face, or the bottom portion of groove in carrier element is placed on, capping is inlaid in carrier element top seal.
9. the carrier for cultured cells according to claim 1, it is characterised in that built-in core thing by it is single it is block,
Sheet or particle are constituted, or are made up of multiple block, sheets or particle, or by powder constituent, can be moved according to magnetic direction.
10. the carrier for cultured cells according to claim 1, it is characterised in that carrier element material uses plastics
Material, non-metallic material, metal or alloying metal material.
11. carriers for cultured cells according to claim 1, it is characterised in that carrier is inserting cell culture appearance
In device and during cultured cells, the size value of carrier should meet x<L/2, wherein x are any path length of carrier), L is in culture vessel
Any one length in two length of sides most short in portion's die cavity, when the size of the length of side can meet loading multiple carrier, its size
Value scope should meet below equation：L/n>x>L/(n+1/2)；Optimal size value scope is；L/(n+1/10)>x>L/(n+1/
3), n is the number of carrier.
12. carriers for cultured cells according to claim 1, it is characterised in that the surface of carrier using hydrophilic or
Hydrophobic treatment, or surface attachment Thermo-sensitive Molecularly Imprinted Polymer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710107175.6A CN106754627A (en) | 2017-02-27 | 2017-02-27 | A kind of carrier for cultured cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710107175.6A CN106754627A (en) | 2017-02-27 | 2017-02-27 | A kind of carrier for cultured cells |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106754627A true CN106754627A (en) | 2017-05-31 |
Family
ID=58959562
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710107175.6A Pending CN106754627A (en) | 2017-02-27 | 2017-02-27 | A kind of carrier for cultured cells |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106754627A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108300713A (en) * | 2017-12-31 | 2018-07-20 | 宁波大学 | The method and device of fixed cell |
| CN110310796A (en) * | 2019-07-28 | 2019-10-08 | 金华市新利磁业工贸有限公司 | A kind of Magnetic force ball and its production method |
| CN111315870A (en) * | 2017-11-09 | 2020-06-19 | 牛津大学科技创新有限公司 | Macro-carriers |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101416059A (en) * | 2003-07-17 | 2009-04-22 | 格洛伯塞尔解决方案公司 | Automated cell culture system and process |
| CN102409020A (en) * | 2010-09-26 | 2012-04-11 | 上海泰因生物技术有限公司 | Non woven/polyester fiber carriers for culturing cells and use method thereof |
| CN103275923A (en) * | 2013-05-30 | 2013-09-04 | 冯玉萍 | GF microcarrier for cell culture and preparation method thereof |
| CN103289948A (en) * | 2013-05-30 | 2013-09-11 | 冯玉萍 | Application of GF (glass fiber) microcarrier for cell culture in adherent dependent cell culture |
| CN103409361A (en) * | 2013-06-24 | 2013-11-27 | 上海瀚正生物技术服务有限公司 | Thermosensitive microcarrier as well as preparation technology and application method thereof |
-
2017
- 2017-02-27 CN CN201710107175.6A patent/CN106754627A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101416059A (en) * | 2003-07-17 | 2009-04-22 | 格洛伯塞尔解决方案公司 | Automated cell culture system and process |
| CN102409020A (en) * | 2010-09-26 | 2012-04-11 | 上海泰因生物技术有限公司 | Non woven/polyester fiber carriers for culturing cells and use method thereof |
| CN103275923A (en) * | 2013-05-30 | 2013-09-04 | 冯玉萍 | GF microcarrier for cell culture and preparation method thereof |
| CN103289948A (en) * | 2013-05-30 | 2013-09-11 | 冯玉萍 | Application of GF (glass fiber) microcarrier for cell culture in adherent dependent cell culture |
| CN103409361A (en) * | 2013-06-24 | 2013-11-27 | 上海瀚正生物技术服务有限公司 | Thermosensitive microcarrier as well as preparation technology and application method thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111315870A (en) * | 2017-11-09 | 2020-06-19 | 牛津大学科技创新有限公司 | Macro-carriers |
| CN108300713A (en) * | 2017-12-31 | 2018-07-20 | 宁波大学 | The method and device of fixed cell |
| CN110310796A (en) * | 2019-07-28 | 2019-10-08 | 金华市新利磁业工贸有限公司 | A kind of Magnetic force ball and its production method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106754627A (en) | A kind of carrier for cultured cells | |
| CN106119186B (en) | It is a kind of for the serum free medium and preparation method thereof for cultivating mdck cell that suspends entirely | |
| CN103119151A (en) | Culture substrate | |
| CN106520552B (en) | A kind of biological reactor for cell culture | |
| CN106701577A (en) | Multifunctional bioreactor used for cell culture and cell sorting | |
| CN103898041B (en) | The cultural method of hybridoma | |
| CN106554937B (en) | A kind of external three-dimensional culture method of liver cell | |
| WO2017080117A1 (en) | Three-dimensional cell culture support and preparation method thereof | |
| CN104687207A (en) | Making method of chlorella drink | |
| CN104694474B (en) | A kind of cell culture processes | |
| CN105121621A (en) | Bioreactor system and methods for alternative cell culture between static and dynamic | |
| CN108699500A (en) | The support fixture and cell culture processes of cell culture container, cell culture container | |
| CN211445767U (en) | Microfluidic chip for three-dimensional culture of cell clusters | |
| CN108728356A (en) | Device and co-culture method for the pairing of different three-dimensional cells group | |
| CN206553537U (en) | A kind of carrier for being used to cultivate cell | |
| CN103255123A (en) | Method for mycelium pellet to form mixed mycelium pellet by adsorbing photosynthetic bacteria | |
| JPH0923785A (en) | Mass culture method for ciliata | |
| CN104630148B (en) | A kind of cell ball in-situ preparation method based on hydrogel microwell plate | |
| TWI245797B (en) | Method of culturing microalgae and device therefor | |
| CN105567626A (en) | Micro-carrier for cell culture and preparation method and application thereof | |
| CN209010531U (en) | A kind of microscope cell cultivation equipment | |
| CN207259522U (en) | Tissue Culture Dish for adherent cell growth | |
| CN208378903U (en) | Increase the cell culture roller bottle of cell culture area | |
| JP2010193846A (en) | Lactic acid fermentation method | |
| CN204644370U (en) | Cell suspension culture device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170531 |
|
| WD01 | Invention patent application deemed withdrawn after publication |