Note: Descriptions are shown in the official language in which they were submitted.
CA 02549777 2006-05-08
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.
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CA 02549777 2006-05-08
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.
CA 02549777 2006-05-08
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.
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CA 02549777 2006-05-08
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
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