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Cell culturing method and device

A cell culture and cell technology, applied in tissue cell/virus culture devices, animal cells, vertebrate cells, etc., can solve the problems of low cell concentration in the medium, hindered cell growth, depletion of medium components, etc., to achieve increased proliferation , the effect of preventing movement

Pending Publication Date: 2020-08-07
TOYO SEIKAN GRP HLDG LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] On the other hand, in cell culture, in general, if the cell concentration in the medium becomes too high accompanying the proliferation of cells, depletion of medium components necessary for proliferation, accumulation of metabolites of the cells themselves, etc. The growth of cells is hindered, which reduces the efficiency of cell proliferation. On the other hand, if the concentration of cells in the medium is too low, the efficiency of cell proliferation and induction of differentiation is also reduced.

Method used

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  • Cell culturing method and device
  • Cell culturing method and device

Examples

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no. 1 Embodiment approach

[0063] Refer below figure 2 and image 3 The cell culture method and apparatus according to the embodiment of the present invention will be described. figure 2 is a schematic cross-sectional view of the cell culture device 100 according to the first embodiment, figure 2 (a) shows a state in which the pressing member is raised, figure 2 (b) shows a state where the pressing member is lowered.

[0064] The cell culture device 100 of the present embodiment includes: a holder 5 on which the cell culture bag 1 is placed, a pressing member 6 having a bottom surface 6a that presses the top surface portion 21a of the upper surface film 21, and a support mechanism that supports the pressing member 6 7.

[0065] It should be noted, figure 2 Figure 2 omits illustration of a pump as a liquid-feeding means for injecting and discharging a medium from the port 3 of the cell culture bag 1 , and a peeling member for peeling the spheroid from the recess 4 of the cell culture bag 1 . I...

no. 2 Embodiment approach

[0090] Next, refer to Figure 6 A second embodiment of the present invention will be described. In the figures, illustration of some of the same components as those of the first embodiment is omitted.

[0091] In the present embodiment, after the sealing step described in the first embodiment, in the peeling step, instead of giving physical stimulation to the concave portion 4 of the cell culture bag from the outside, Figure 6 As shown in (a), each recess 4 remains in a closed state, and the cell culture bag 1 is placed in each figure 1 The cell culture device 100 is shown turned upside down.

[0092] Alternatively, you can Figure 6 As shown in (b), in the peeling process, instead of giving physical stimulation to the concave portion from the outside, each concave portion 4 is kept in a closed state, and the cell culture bag 1 is placed one by one. figure 1 The cell culture device 100 is shown tilted. It should be noted that the inclination angle of the cell culture b...

no. 3 Embodiment approach

[0099] Next, refer to Figure 7 A third embodiment of the present invention will be described. In the figures, illustration of some of the same components as those of the first embodiment is omitted.

[0100] In the present embodiment, after the sealing step described in the first embodiment, in the peeling step, instead of vibrating or shaking the recessed part 4 of the cell culture bag, the upper part respectively arranged under the plurality of recessed parts 4 is used. The top member 9 pushes up each concave portion from below to give physical stimulation to each concave portion 4 .

[0101] The top member 9 moves up and down by an actuator (not shown) in the opening 5b formed on the loading surface 5a of the stand 5 . The bottom portion of the crown-shaped concave portion 4 that is convexly curved downward is pushed upward by the upper top member 9 , and bulges reversely so as to be convexly curved upward. By pushing the concave portion 4 upward, the spherical body C a...

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Abstract

The present invention provides a cell culturing device that, when culturing cells using a cell culture bag in which a plurality of recesses is formed, releases cells or cell aggregates adhering to theinner surfaces of the recesses while preventing movement of cells between the recesses, thereby improving the proliferation / differentiation induction efficiency of cells or cell aggregates. The present invention is a cell culturing method that uses a cell culturing bag (1) that comprises a bag body (2) configured from an upper surface film (21) and a lower surface film (22) having a sealed periphery, and a port (3) attached to the bag body (2), a plurality of recesses (4) being formed in the lower surface film (22), and the method comprising: a closing-off step for discharging a culture medium (S) contained in the bag body (2) through the port (3), and closing off the plurality of recesses (4) using the upper surface film (21); and a releasing step for releasing, in some or all of the plurality of closed-off recesses (4), spheroids adhering to the inner surfaces of the recesses (4).

Description

technical field [0001] The present invention relates to a cell culture method and device, and more specifically, to a cell culture method and device using a cell culture bag provided with a plurality of recesses. Background technique [0002] In recent years, in the fields of drug production, gene therapy, regenerative medicine, and immunotherapy, cells (including tissues, microorganisms, viruses, etc.) are required to be efficiently mass-cultured and induced to differentiate in an artificial environment. [0003] In such cell culture, in order to avoid the risk of contamination, a closed system cell culture bag is sometimes used. The cell culture bag is composed of, for example, a bag body made of a plastic film that seals the periphery, and a port attached to the bag body. Patent Document 1 describes a cell culture bag that suppresses the movement of cells in the cell culture bag by providing a plurality of recesses on the bottom surface of the bag body. [0004] On the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00C12M1/00C12M3/00C12N5/071
CPCC12M23/14C12M23/12C12M33/00C12M41/40C12M27/16C12M23/50C12M29/26C12N5/0068C12N2527/00C12N2513/00
Inventor 末永亮光石真稀
Owner TOYO SEIKAN GRP HLDG LTD
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