Cell culture device and movement system

Patent metadata

Field	Value
Publication number	US-2024368517-A1
Application number	US-202218580711-A
Country	US
Kind code	A1
Filing date	Jul 19, 2022
Priority date	Jul 19, 2021
Publication date	Nov 7, 2024
Grant date	—

Abstract

A cell culture apparatus including a cell culture device and a movement system for moving the cell culture device. The cell culture device includes at least two reservoirs for holding a liquid cell medium, one or more chambers for culturing of living cells, tissues or living organoids and at least two perfusion channels connecting the reservoirs. The chambers are separated from at least one of the perfusion channels by a semipermeable barrier for selective transport of cell media and/or for selective growth or migration of living cells. The movement system is configured to move the cell culture device with a movement axis in a tilted orientation in order to move the lowest point of a flow loop through all points within the flow path of the flow loop and thereby generate gravity driven circulation of the liquid cell medium around the flow loop.

First claim

We claim:

1 . A cell culture apparatus comprising: a cell culture device; and a movement system for moving the cell culture device;
the cell culture device comprising: at least two reservoirs for holding a liquid cell medium, one or more chambers for culturing of living cells, tissues or living organoids and at least two perfusion channels connecting the reservoirs; wherein the one or more chambers are separated from at least one of the perfusion channels by a semipermeable barrier for selective transport of cell media and/or for selective growth or migration of living cells; and wherein the reservoirs and perfusion channels are in fluid communication with one another in order to form a flow loop having a flow path for a one-way gravity-driven flow of the liquid cell medium when the cell culture device is tilted and moved by the movement system; a movement axis being defined as an axis of the cell culture device that passes through the cell culture device with the flow loop being located around the movement axis; and the movement system being configured to move the cell culture device with the movement axis in a tilted orientation in order to move the lowest point of the flow loop through all points within the flow path of the flow loop and thereby generate gravity driven circulation of the liquid cell medium around the flow loop, wherein the tilted orientation places the movement axis at an angle in the range 5° to 85° away from the vertical.

2 . A cell culture apparatus as claimed in claim 1 , wherein the movement in a tilted orientation is a movement of the device whilst the axis is at an angle of 20° to 80° away from the vertical.

3 . A cell culture apparatus as claimed in claim 1 , wherein the barrier acts as a semi-permeable and/or selectively permeable barrier between the perfusion channel and the chamber, allowing for passage of certain cell media to provide a way of connecting the liquid cell medium to the at least one chamber.

4 . A cell culture apparatus as claimed in claim 1 , wherein the barrier is an extra cellular matrix (ECM) barrier, and/or wherein the barrier includes a biomaterial or extracellular natural material and comprises one or more of Matrigel®, Geltrex®, Collagen, a synthetic hydrogel, and/or a polymer membrane.

5 . (canceled)

6 . A cell culture apparatus as claimed in claim 1 , comprising two barriers at two locations about the chamber, wherein the two barriers each provide for transport of certain cell media between the chamber and the two perfusion channels.

7 . A cell culture apparatus as claimed in claim 1 , wherein a part of the flow loop is configured to allow contact of the liquid cell medium with a gas from outside of the flow loop.

8 . A cell culture apparatus as claimed in claim 1 , the movement axis for the cell culture device being defined as an axis normal to a plane of the device, wherein during movement of the device by the movement system the movement axis is tilted in the sense that it is angled to the vertical thereby positioning lifting one section of the flow loop higher than another section of the flow loop, and wherein the position of the tilted axis moves in order to change the section(s) of the flow loop that are higher and lower relative to one another.

9 . A cell culture apparatus as claimed in claim 1 , wherein the tilted movement axis is moved by the movement system in a rotational fashion so that a reference point on the movement axis traces around the circumference of a two dimensional shape.

10 . A cell culture apparatus as claimed in claim 1 , wherein the movement in a tilted orientation includes a rotation of the cell culture device about the movement axis, such that a reference point on the flow loop moves in a circle about the tilted movement axis.

11 . A cell culture apparatus as claimed in claim 1 , wherein the flow path includes a flow reversal restriction feature comprising one or more of: a change in depth; a capillary stop valve; and/or a surface treatment promoting decreased wetting.

12 . A cell culture apparatus as claimed in claim 1 , wherein the flow loop comprises a looped flow path that passes, in sequence, from a first reservoir, then through a first perfusion channel and past a first barrier at a first side of the chamber, then to a second reservoir, then from the second reservoir, through a second perfusion channel and past a second barrier at a second side of the chamber, then returning to the first reservoir.

13 . A cell culture apparatus as claimed in claim 1 , wherein the reservoirs each comprise a volume of 40 to 900 μL for receiving liquid cell medium.

14 . A cell culture apparatus as claimed in claim 1 , wherein the perfusion channels each extend from the outlet of one reservoir to the inlet of another reservoir, passing by at least one chamber with a barrier providing a link to the chamber at a mid-portion of the channel; and wherein the length of each perfusion channel is in the range 10 to 30 mm.

15 . A cell culture apparatus as claimed in claim 1 , wherein the chamber comprises a volume or 2.5 to 120 μL for receiving the living cells or living organoids and the chamber is protected from shear force of the perfusion channels via the barrier(s).

16 . A cell culture apparatus as claimed in claim 1 , wherein the cell culture device includes the liquid cell medium therein and/or the chamber includes therein a living cell or living organoid.

17 . A cell culture apparatus as claimed in claim 16 , wherein the cell culture device includes a volume of 50 to 600 μL of the liquid cell medium.

18 . A cell culture apparatus as claimed in claim 1 , wherein the dimensions of the cell culture device as well as the tilting speed and angle of movement by the movement system are configured to provide flow rates within the range of physiological flow rates such that the apparatus is configured to provide a flow rate of 15 to 80 μL/s of the liquid cell medium in the flow path.

19 . (canceled)

20 . A cell culture apparatus as claimed in claim 1 , wherein the dimensions of the cell culture device as well as the tilting speed and angle of movement by the movement system are configured to provide shear stresses within the range of physiological shear stresses such that the apparatus is configured to provide a shear stress in the range of 0.1 to 100 dyne/cm 2 for flow of the liquid cell medium in the flow path.

21 . (canceled)

22 . A cell culture device for the cell culture apparatus of claim 1 , the cell culture device comprising:
at least two reservoirs for holding a liquid cell medium, one or more chambers for culturing of living cells or living organoids and at least two perfusion channels connecting the reservoirs; wherein the one or more chambers are separated from at least one of the perfusion channels by a semipermeable barrier for selective transport of cell media and/or for selective growth or migration of living cells; and wherein the reservoirs and perfusion channels are in fluid communication with one another in order to form a flow loop having a flow path for a one-way gravity-driven flow of the liquid cell medium when the cell culture device is tilted and moved by the movement system; wherein a movement axis is defined as an axis of the cell culture device that passes through the cell culture device with the flow loop being located around the movement axis; and wherein the cell culture device is configured such that when it is moved with the movement axis in a tilted o

Assignees

• Univ Oslo
• Univ Oslo Hf

Inventors

• Krauss Stefan Johannes Karl
• Busek Mathias
• Martinez Mikel Amirola
• Aizenshtadt Aleksandra

Classifications

• C12M29/10

  Perfusion · CPC title

• C12M29/04

  Filters; Permeable or porous membranes or plates, e.g. dialysis · CPC title

• C12M23/24

  Gas permeable parts · CPC title

• C12M21/08

  for producing artificial tissue or for ex-vivo cultivation of tissue (prostheses A61F2/00, grafts A61L27/00) · CPC title

• C12M27/16

  Vibrating; Shaking; Tilting · CPC title