CA2549777A1 - A novel assembly for passaging cells - Google Patents
A novel assembly for passaging cells Download PDFInfo
- Publication number
- CA2549777A1 CA2549777A1 CA002549777A CA2549777A CA2549777A1 CA 2549777 A1 CA2549777 A1 CA 2549777A1 CA 002549777 A CA002549777 A CA 002549777A CA 2549777 A CA2549777 A CA 2549777A CA 2549777 A1 CA2549777 A1 CA 2549777A1
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- Prior art keywords
- component
- holes
- cell culture
- modular assembly
- assembly
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- 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.)
- Abandoned
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- C—CHEMISTRY; METALLURGY
- 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
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/06—Plates; Walls; Drawers; Multilayer plates
-
- C—CHEMISTRY; METALLURGY
- 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
- C12M23/10—Petri dish
-
- C—CHEMISTRY; METALLURGY
- 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
- C12M23/38—Caps; Covers; Plugs; Pouring means
-
- C—CHEMISTRY; METALLURGY
- 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
- C12M23/44—Multiple separable units; Modules
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- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Clinical Laboratory Science (AREA)
- Immunology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention relates generally to laboratory ware, and more particularly to vessels, such as flasks and dishes, and methods for cultivating and passaging biological cell cultures therein. Cell culture vessels are disclosed providing a means of trypsin-free passaging of adherent cell cultures. A
representative embodiment of the invention, consisting of interlocking stackable culture dishes which allow for the separation of the growth surface into fractions, is disclosed.
representative embodiment of the invention, consisting of interlocking stackable culture dishes which allow for the separation of the growth surface into fractions, is disclosed.
Description
DESCRIPTIDN
Field of the invention The invention relates generally to laboratory ware, and more particularly to vessels, such as flasks and dishes, and methods for cultivating and passaging biological cell cultures therein.
Description of prior art This invention relates to an article to be used for cultivating and passaging adherent biological cells without trypsinization. Passaging herein refers to the separation of a population of cells growing on a solid surface into subpopulations of given size. Prior art in passaging adherent cells involves trypsinization of the parent cell culture followed by re-plating at known cell densities.
Trypsinization involves the use of the protease trypsin in solution which degrades extracellular proteins (such as adhesins and extracellular polymeric matrix) to separate individual cells from each other and from the solid surface upon which they grow. The trypsin is then removed or neutralized and cells are separated into subpopulations and allowed to re-attach to new growth surfaces.
Passaging cells is required to expand a population for experimentation or maintain its density despite cell division. Trypsinization is time consuming, requires specialized technical knowledge and is hard to automate. Certain biological downsides to trypsinization also include, but are not restricted to, proteolytic damage to cells, selection of cells that survive proteolytic damage, selection of cells that adhere following trypsinization and selection of cells that adhere poorly prior to trypsinization. No prior art discloses a method of passaging cells mechanically.
Object of the invention Our assembly allows for the passaging of adherent cell culture populations into subpopulations without selection, without the use of trypsin and in a manner that easily allows for automation.
These and other objects are achieved by embodiments of the invention described herein.
manner that will create a coplanar surface with the bottom of the upper component. Said bottom component being shaped to accommodate central component and shaped to fit within the cover component.
The central component shaped to fit within the bottom component and shaped to accommodate upper component. Said component having holes in its bottom (lE) shaped and aligned to tightly accommodate elevated surfaces (1F) from the bottom component and having a fraction (1D) of its bottom surface elevated, shaped and aligned to completely fill holes in the bottom of the upper (1C) component in a manner that will create a coplanar surface with the bottom of the upper component.
Said elevated fraction creating an unbroken coplanar surface when coupled with bottom and upper components. Said central component having a means (1B) by which automated devices could manipulate said component in this embodiment.
The upper component shaped to fit within the central component. Said component having holes in its bottom (1C) shaped to tightly accommodate elevated surfaces (1F) from the bottom component and elevated surfaces (1D) from the central component. Said holes aligned and shaped in a manned that will create an unbroken coplanar surface when coupled with bottom and central components. Said central component having a means (1B) by which automated devices could manipulate said component.
The cover component shaped to completely block vertical transfer above the media retained by the walls encircling the coplanar surface created by the assembled upper, central and bottom components. Said cover component having a means to be retained above assembled upper, central and bottom components, such as, but not restricted to, walls extending down to encircle bottom component. Said cover having a means (1A) by which automated devices could manipulate said cover.
It may be desirable to reduce the surface area of the elevated surfaces ID and 1F while increasing their number to reduce the total number of cells that can adhere to the elevated surface.
Summary of the invention In a particularly advantageous embodiment of the illustrated invention, a modular assembly of three stackable components consists of a system of flat-top pins and holes.
The surface of the upper component contains a number of evenly distributed holes, the central component contains an array of evenly distributed pins which fit into half the holes of the upper component and an array of holes aligned with the remaining holes of the upper component, and the bottom component contains an array of longer pins which fit into the holes of the central component and the remaining holes of the upper component in a manner that creates an unbroken co-planar surface upon which cells can grow.
The particular advantage of this system is the lack of need for trypsinization to separate cells into subpopulations of given size, and the potential for easy automation.
Adherent cells can be separated into three subpopulations (in this particular embodiment): cells growing on the surface of the flat-top pins of the bottom and central components and cells growing on the planar surface of the upper component.
Each component with cells on its surface can be used to seed a new assembly formed with the two other components. This judgement-free method of separating cells is particularly well suited for automation, removing the need for time consuming and labour intensive maintenance of cell cultures.
Description of the drawings Fig. 1 is a side view of a modular cell culture assembly embodying the disclosed invention.
Fig. 2 is a side perspective view of the modular cell culture assembly shown in Fig. 1.
Fig. 3A is a side perspective view of the bottommost module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig. 3B is a top view of the bottommost module of the modular cell culture assembly show in Fig. 1.
Fig. 3C is a side view of the bottommost module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 4A is a top perspective view of the central module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 4B is a bottom perspective view of the central module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Field of the invention The invention relates generally to laboratory ware, and more particularly to vessels, such as flasks and dishes, and methods for cultivating and passaging biological cell cultures therein.
Description of prior art This invention relates to an article to be used for cultivating and passaging adherent biological cells without trypsinization. Passaging herein refers to the separation of a population of cells growing on a solid surface into subpopulations of given size. Prior art in passaging adherent cells involves trypsinization of the parent cell culture followed by re-plating at known cell densities.
Trypsinization involves the use of the protease trypsin in solution which degrades extracellular proteins (such as adhesins and extracellular polymeric matrix) to separate individual cells from each other and from the solid surface upon which they grow. The trypsin is then removed or neutralized and cells are separated into subpopulations and allowed to re-attach to new growth surfaces.
Passaging cells is required to expand a population for experimentation or maintain its density despite cell division. Trypsinization is time consuming, requires specialized technical knowledge and is hard to automate. Certain biological downsides to trypsinization also include, but are not restricted to, proteolytic damage to cells, selection of cells that survive proteolytic damage, selection of cells that adhere following trypsinization and selection of cells that adhere poorly prior to trypsinization. No prior art discloses a method of passaging cells mechanically.
Object of the invention Our assembly allows for the passaging of adherent cell culture populations into subpopulations without selection, without the use of trypsin and in a manner that easily allows for automation.
These and other objects are achieved by embodiments of the invention described herein.
manner that will create a coplanar surface with the bottom of the upper component. Said bottom component being shaped to accommodate central component and shaped to fit within the cover component.
The central component shaped to fit within the bottom component and shaped to accommodate upper component. Said component having holes in its bottom (lE) shaped and aligned to tightly accommodate elevated surfaces (1F) from the bottom component and having a fraction (1D) of its bottom surface elevated, shaped and aligned to completely fill holes in the bottom of the upper (1C) component in a manner that will create a coplanar surface with the bottom of the upper component.
Said elevated fraction creating an unbroken coplanar surface when coupled with bottom and upper components. Said central component having a means (1B) by which automated devices could manipulate said component in this embodiment.
The upper component shaped to fit within the central component. Said component having holes in its bottom (1C) shaped to tightly accommodate elevated surfaces (1F) from the bottom component and elevated surfaces (1D) from the central component. Said holes aligned and shaped in a manned that will create an unbroken coplanar surface when coupled with bottom and central components. Said central component having a means (1B) by which automated devices could manipulate said component.
The cover component shaped to completely block vertical transfer above the media retained by the walls encircling the coplanar surface created by the assembled upper, central and bottom components. Said cover component having a means to be retained above assembled upper, central and bottom components, such as, but not restricted to, walls extending down to encircle bottom component. Said cover having a means (1A) by which automated devices could manipulate said cover.
It may be desirable to reduce the surface area of the elevated surfaces ID and 1F while increasing their number to reduce the total number of cells that can adhere to the elevated surface.
Summary of the invention In a particularly advantageous embodiment of the illustrated invention, a modular assembly of three stackable components consists of a system of flat-top pins and holes.
The surface of the upper component contains a number of evenly distributed holes, the central component contains an array of evenly distributed pins which fit into half the holes of the upper component and an array of holes aligned with the remaining holes of the upper component, and the bottom component contains an array of longer pins which fit into the holes of the central component and the remaining holes of the upper component in a manner that creates an unbroken co-planar surface upon which cells can grow.
The particular advantage of this system is the lack of need for trypsinization to separate cells into subpopulations of given size, and the potential for easy automation.
Adherent cells can be separated into three subpopulations (in this particular embodiment): cells growing on the surface of the flat-top pins of the bottom and central components and cells growing on the planar surface of the upper component.
Each component with cells on its surface can be used to seed a new assembly formed with the two other components. This judgement-free method of separating cells is particularly well suited for automation, removing the need for time consuming and labour intensive maintenance of cell cultures.
Description of the drawings Fig. 1 is a side view of a modular cell culture assembly embodying the disclosed invention.
Fig. 2 is a side perspective view of the modular cell culture assembly shown in Fig. 1.
Fig. 3A is a side perspective view of the bottommost module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig. 3B is a top view of the bottommost module of the modular cell culture assembly show in Fig. 1.
Fig. 3C is a side view of the bottommost module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 4A is a top perspective view of the central module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 4B is a bottom perspective view of the central module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 4C is a top view of the central module of the modular cell culture assembly show in Fig. 1.
Fig 4D is a side view of the central module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 5A is a bottom perspective view of the upper module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 5B is a top view of the upper module of the modular cell culture assembly show in Fig. 1.
Fig 5C is a side view of the upper module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 6A is a top perspective view of the cover of the modular cell culture assembly show in Fig.
1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 6B is a bottom perspective view of the cover of the modular cell culture assembly show in Fig. 1.
Fig 6C is a top view of the cover of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 6D is a top perspective view of the cover of the modular cell culture assembly show in Fig.
1.
Fig 7A is a top perspective view of the assembled modular cell culture assembly show in Fig.
1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 7B is a bottom perspective view of the assembled modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 7C is a top view of the assembled modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 7D is a side view of the assembled modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Detailed description of the drawings Referring now to figures 1 and 2. A cell culture assembly, comprised of 3 interlocking components (a bottom component (figure 3), a central component (figure 4), an upper component (figure 5)) and a cover; the bottom, central and upper components creating an unbroken coplanar surface once assembled (as in Fig 7) and having continuous walls able to retain liquid media on the coplanar surface in this embodiment.
The bottom component having a fraction (1F) of its bottom surface elevated, shaped and aligned to completely fill holes in the bottom of the central (lE) and upper (1C) components in a
Fig 4D is a side view of the central module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 5A is a bottom perspective view of the upper module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 5B is a top view of the upper module of the modular cell culture assembly show in Fig. 1.
Fig 5C is a side view of the upper module of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 6A is a top perspective view of the cover of the modular cell culture assembly show in Fig.
1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 6B is a bottom perspective view of the cover of the modular cell culture assembly show in Fig. 1.
Fig 6C is a top view of the cover of the modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 6D is a top perspective view of the cover of the modular cell culture assembly show in Fig.
1.
Fig 7A is a top perspective view of the assembled modular cell culture assembly show in Fig.
1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 7B is a bottom perspective view of the assembled modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 7C is a top view of the assembled modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Fig 7D is a side view of the assembled modular cell culture assembly show in Fig. 1, with transparency to reveal features hidden by the geometry of the assembly.
Detailed description of the drawings Referring now to figures 1 and 2. A cell culture assembly, comprised of 3 interlocking components (a bottom component (figure 3), a central component (figure 4), an upper component (figure 5)) and a cover; the bottom, central and upper components creating an unbroken coplanar surface once assembled (as in Fig 7) and having continuous walls able to retain liquid media on the coplanar surface in this embodiment.
The bottom component having a fraction (1F) of its bottom surface elevated, shaped and aligned to completely fill holes in the bottom of the central (lE) and upper (1C) components in a
Claims (8)
1. A modular assembly for cultivating and passaging adherent cell cultures consisting of a total cell culture area comprising of a set of at least two interlocking co-planar surfaces configured and sized to be spannable by biological cells.
2. The modular assembly of claim 1, wherein each individual component comprises a fraction of the total cell culture area.
3. The modular assembly of claim 1, wherein the coplanar surfaces of each component is adequate for adherent cell culture.
4. The modular assembly of claim 1, wherein each coplanar surface is divided into multiple smaller units which are evenly distributed over the cell culture area.
5. The modular assembly of claim 1, wherein the interlocking components are stackable.
6. The modular assembly of claim 1, wherein the system of interlocking coplanar surfaces consist of a system of flat-top pins and holes.
7. The modular assembly of claim 1, wherein the system of flat-top pins and holes consists of three stackable components: the surface of the upper component containing a number of evenly distributed holes, the central component containing an array of evenly distributed pins which fit into half the holes of the upper component and an array of holes aligned with the remaining holes of the upper component, and the bottom component containing an array of longer pins which fit into the holes of the central component and the remaining holes of the upper component.
8. The modular assembly of claim 1 including a means of sterility such as a cover component.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA002549777A CA2549777A1 (en) | 2006-05-08 | 2006-05-08 | A novel assembly for passaging cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA002549777A CA2549777A1 (en) | 2006-05-08 | 2006-05-08 | A novel assembly for passaging cells |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2549777A1 true CA2549777A1 (en) | 2007-11-08 |
Family
ID=38663479
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002549777A Abandoned CA2549777A1 (en) | 2006-05-08 | 2006-05-08 | A novel assembly for passaging cells |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2549777A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015121302A1 (en) * | 2014-02-12 | 2015-08-20 | Ekeroth Kristian | Dividable surfaces for cell culturing |
-
2006
- 2006-05-08 CA CA002549777A patent/CA2549777A1/en not_active Abandoned
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015121302A1 (en) * | 2014-02-12 | 2015-08-20 | Ekeroth Kristian | Dividable surfaces for cell culturing |
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| FZDE | Dead |