JP2020174574A5

JP2020174574A5 -

Info

Publication number: JP2020174574A5
Application number: JP2019078996A
Authority: JP
Filing date: 2019-04-18
Publication date: 2022-03-04

Description

上記目的を達成するために、本発明は以下の手段を提供する。
本発明の第１態様は、培養容器内の特定の領域の培地に含まれている細胞の数または細胞密度を計測する観察部と、前記特定の領域の培地を排出する排出部と、前記観察部によって計測された前記特定の領域の培地に含まれている前記細胞の数または細胞密度が所定の閾値よりも少ない場合に前記排出部によって前記特定の領域の培地を排出させ、前記観察部によって計測された前記特定の領域の培地に含まれている前記細胞の数または細胞密度が所定の閾値以上の場合は前記排出部による前記特定の領域の培地の排出を行わない制御部とを備える培地交換システムである。 In order to achieve the above object, the present invention provides the following means.
The first aspect of the present invention is an observation unit for measuring the number or cell density of cells contained in a medium in a specific region in a culture vessel, a discharge unit for discharging the medium in the specific region, and the observation. When the number or cell density of the cells contained in the medium of the specific region measured by the unit is less than a predetermined threshold value, the medium of the specific region is discharged by the discharge unit, and the medium of the specific region is discharged by the observation unit. A medium provided with a control unit that does not discharge the medium of the specific region by the discharge unit when the number or cell density of the cells contained in the measured medium of the specific region is equal to or higher than a predetermined threshold value. It is an exchange system.

上記態様に係る培地交換システムは、前記培養容器から前記特定の領域の培地を吸引する吸引部と、該吸引部によって吸引された前記培地を前記培養容器に返送する返送部とを備え、前記観察部が、前記吸引部によって吸引された前記培地に含まれている前記細胞の数または細胞密度を計測し、前記制御部が、前記観察部によって計測された前記細胞の数または細胞密度が所定の閾値よりも少ない場合は、前記吸引部によって吸引された前記培地を前記排出部によって排出させ、前記観察部によって計測された前記細胞の数または細胞密度が所定の閾値以上の場合は、前記吸引部によって吸引された前記培地を前記返送部によって前記培養容器に返送させることとしてもよい。 The culture medium exchange system according to the above aspect includes a suction unit that sucks the medium of the specific region from the culture container and a return unit that returns the culture medium sucked by the suction unit to the culture container. The unit measures the number or cell density of the cells contained in the medium sucked by the suction unit, and the control unit measures the number or cell density of the cells measured by the observation unit. If it is less than the threshold value, the medium sucked by the suction unit is discharged by the discharge unit, and if the number or cell density of the cells measured by the observation unit is equal to or higher than a predetermined threshold value, the suction unit is used. The medium sucked by the medium may be returned to the culture vessel by the return unit.

上記態様に係る培地交換システムは、前記観察部によって前記細胞の数または細胞密度が計測された前記培地に対して前記排出部によって排出する流路と前記返送部によって返送する流路とを切り換える切換機構を備え、前記制御部が、前記切換機構による前記流路の切り換えを制御することとしてもよい。
この構成により、制御部によって切り換え機構を制御するだけの簡易な構成で、観察部によって細胞の数が計数された培地をそのまま排出するかそれとも培養容器に戻すかの切り換える行うことができる。 The medium exchange system according to the above aspect switches between a flow path discharged by the discharging unit and a flow path returned by the returning unit with respect to the medium for which the number of cells or the cell density has been measured by the observation unit. A mechanism may be provided, and the control unit may control switching of the flow path by the switching mechanism.
With this configuration, it is possible to switch between discharging the medium in which the number of cells has been counted by the observation unit as it is or returning it to the culture vessel, with a simple configuration in which the switching mechanism is only controlled by the control unit.

上記態様に係る培地交換システムは、前記吸引部によって吸引された前記培地の体積および／または前記吸引部によって吸引された前記培地に含まれる前記細胞の数または細胞密度を記録する記録部を備えることとしてもよい。
この構成により、記録部に記録された培地の体積やその培地中に含まれる細胞数を用いて、培地交換の質を評価することができる。 The medium exchange system according to the above aspect includes a recording unit that records the volume of the medium sucked by the suction unit and / or the number or cell density of the cells contained in the medium sucked by the suction unit. May be.
With this configuration, the quality of medium exchange can be evaluated using the volume of the medium recorded in the recording unit and the number of cells contained in the medium.

本発明の第２態様は、培養容器内の特定の領域の培地に含まれている細胞の数または細胞密度を取得する細胞数取得ステップと、該細胞数取得ステップによって取得された前記細胞の数または細胞密度が所定の閾値と比較して少ないか否かを判定する判定ステップと、該判定ステップによって前記細胞の数または細胞密度が前記所定の閾値よりも少ないと判定された場合にのみ前記特定の領域の培地を排出する培地処理ステップとを含む培地交換方法である。 A second aspect of the present invention is a cell number acquisition step for acquiring the number or cell density of cells contained in a medium in a specific region in a culture vessel, and the number of cells acquired by the cell number acquisition step. Alternatively, the specification is performed only when a determination step for determining whether or not the cell density is lower than the predetermined threshold and when the determination step determines that the number of cells or the cell density is less than the predetermined threshold. It is a culture medium exchange method including a culture medium treatment step of draining the culture medium of the region.

上記態様に係る培地交換方法は、前記培養容器から前記特定の領域の培地を吸引する吸引ステップを含み、前記細胞数取得ステップが、前記吸引ステップによって吸引された前記培地に含まれている前記細胞の数または細胞密度を取得し、前記培地処理ステップが、前記判定ステップによって前記細胞の数または細胞密度が前記所定の閾値よりも少ないと判定された場合は、前記吸引ステップによって吸引された前記培地を排出し、前記判定ステップによって前記細胞の数または細胞密度が前記所定の閾値以上と判定された場合は、前記吸引ステップによって吸引された前記培地を前記培養容器に返送することとしてもよい。 The medium exchange method according to the above embodiment includes a suction step of sucking the medium of the specific region from the culture vessel, and the cell number acquisition step is the cells contained in the medium sucked by the suction step. If the number or cell density of the cells is determined by the determination step to be less than the predetermined threshold, the medium sucked by the suction step is obtained. When the number of cells or the cell density is determined to be equal to or higher than the predetermined threshold value by the determination step, the medium sucked by the suction step may be returned to the culture vessel.

観察部７としては、細胞Ｓを非染色かつ非標識で観察する既存の技術を採用できる。観察部７は、例えば、培養容器３内の培地Ｗおよび細胞Ｓが存在する任意の領域である特定領域Ｒに培養容器３の外部から照明光を照射するＬＥＤ（Light Emitting Diode）、レーザ光等の光源と、照明光が照射された特定領域Ｒからの観察光を培養容器３の外部において集光するレンズと、レンズによって集光された観察光を撮影することにより特定領域Ｒの画像を取得するＣＣＤイメージセンサまたはＣＭＯＳイメージセンサ等の撮像素子と、撮像素子によって取得された画像を処理する画像処理部（いずれも図示略）とを備えている。 As the observation unit 7, an existing technique for observing cells S unstained and unlabeled can be adopted. The observation unit 7 is, for example, an LED (Light Emitting Diode), a laser beam, or the like that irradiates a specific region R, which is an arbitrary region in the culture vessel 3 in which the medium W and the cells S are present, with illumination light from the outside of the culture vessel 3. The image of the specific area R is acquired by photographing the light source of the above, a lens that collects the observation light from the specific area R irradiated with the illumination light outside the culture vessel 3, and the observation light collected by the lens. It includes an image pickup element such as a CCD image sensor or a CMOS image sensor, and an image processing unit (both not shown) for processing an image acquired by the image pickup element.

Claims

An observation unit that measures the number or density of cells contained in the medium of a specific area in the culture vessel,
A discharge section that discharges the medium in the specific region, and a discharge section.
When the number or cell density of the cells contained in the medium of the specific region measured by the observation unit is less than a predetermined threshold value, the discharge unit discharges the medium of the specific region, and the observation. When the number or cell density of the cells contained in the medium of the specific region measured by the unit is equal to or higher than a predetermined threshold value, the discharge unit does not discharge the medium of the specific region. A medium exchange system equipped.

The culture medium exchange system according to claim 1, wherein the discharge unit includes a discharge channel member for circulating the medium and a discharge pump for transferring the medium to the outside of the culture vessel via the drainage channel member. ..

A suction unit that sucks the medium of the specific region from the culture vessel,
It is provided with a return unit for returning the medium sucked by the suction unit to the culture container.
The observation unit measures the number or cell density of the cells contained in the medium sucked by the suction unit.
When the control unit discharges the medium sucked by the suction unit by the discharge unit when the number of cells or the cell density measured by the observation unit is less than a predetermined threshold value, the observation unit discharges the medium. The medium according to claim 1 or 2, wherein when the measured number of cells or cell density is equal to or higher than a predetermined threshold value, the medium sucked by the suction unit is returned to the culture vessel by the return unit. Exchange system.

The culture medium exchange system according to claim 3, wherein the return unit includes a return flow path member for circulating the medium and a return pump for transferring the medium to the culture vessel via the return flow path member.

A switching mechanism for switching between a flow path discharged by the discharging section and a flow path returned by the returning section with respect to the medium for which the number or cell density of the cells has been measured by the observation section is provided.
The culture medium exchange system according to claim 3 or 4, wherein the control unit controls switching of the flow path by the switching mechanism.

Claims 3 to 5 include a recording unit that records the volume of the medium sucked by the suction unit and / or the number or cell density of the cells contained in the medium sucked by the suction unit. The medium exchange system according to any one.

A stirring unit that stirs the medium by rotating around a predetermined axis of rotation in the culture vessel, and a stirring unit.
The culture medium exchange system according to any one of claims 1 to 5, further comprising a recording unit for recording the rotation speed of the stirring unit.

The culture medium exchange system according to any one of claims 1 to 5, further comprising a recording unit for recording the pH of the culture medium in the culture vessel.

The discharge section has a suction port for sucking the medium of the specific region in the culture vessel.
The culture medium exchange system according to claim 1 or 2, further comprising an adjusting mechanism capable of adjusting the position of the suction port in the culture vessel.

The suction portion has a suction port for sucking the medium of the specific region in the culture vessel.
The culture medium exchange system according to any one of claims 3 to 5, further comprising an adjusting mechanism capable of adjusting the position of the suction port in the culture vessel.

The culture medium exchange system according to claim 9 or 10, further comprising a recording unit that records the height from the bottom surface of the culture vessel to the suction port.

The culture medium exchange system according to any one of claims 1 to 11, further comprising a culture medium supply unit for supplying the culture medium to the culture vessel.

The culture medium exchange system according to claim 12, wherein the culture medium supply unit supplies the culture medium with the amount of culture medium discharged as a threshold value.

The culture medium exchange system according to any one of claims 1 to 13, wherein the observation unit measures the number of the cells.

The cell number acquisition step to acquire the number of cells or cell density contained in the medium of a specific region in the culture vessel, and
A determination step for determining whether or not the number or cell density of the cells acquired by the cell number acquisition step is smaller than a predetermined threshold value.
A medium exchange method comprising a medium treatment step in which the medium in the specific region is discharged only when the number of cells or the cell density is determined to be less than the predetermined threshold value by the determination step.

Including a suction step of sucking the medium of the particular region from the culture vessel.
The cell number acquisition step acquires the number or cell density of the cells contained in the medium sucked by the suction step.
When the determination step determines that the number or cell density of the cells is less than the predetermined threshold value, the medium sucked by the suction step is discharged, and the determination step determines that the medium is aspirated. The medium exchange method according to claim 15 , wherein when the number of cells or the cell density is determined to be equal to or higher than the predetermined threshold value, the medium sucked by the suction step is returned to the culture vessel.