Cell concentration adjustment device, and automatic subculture system using same

Patent metadata

Field	Value
Publication number	US-10138456-B2
Application number	US-201515327208-A
Country	US
Kind code	B2
Filing date	Jul 8, 2015
Priority date	Jul 22, 2014
Publication date	Nov 27, 2018
Grant date	Nov 27, 2018

Abstract

A device which automatically performs a step in which expanded and cultured cells are diluted to a desired cell concentration and re-inoculated using a cell-concentration adjustment device having an inlet for taking in a cell suspension; an outlet for discharging a diluted cell suspension; and a flow path which is provided between the inlet and the outlet and is capable of holding a cell suspension, the flow path being provided with: a liquid delivery pump for causing a cell suspension inside to flow; a cell-concentration measurement instrument for collecting data related to a cell concentration per unit amount of the cell suspension; and a dilution-liquid container for holding a dilution liquid which is supplied to the flow path to dilute the cell suspension. The device further includes a control unit for controlling at least the liquid delivery pump on the basis of the data obtained by the cell-concentration measurement instrument, wherein the control unit determines, on the basis of the data obtained by the cell-concentration measurement instrument, an amount of the dilution liquid required to bring the cell concentration to the desired concentration, and drives the liquid delivery pump so as to take in the required amount of the dilution liquid into the flow path and mix the cell suspension and the dilution liquid.

First claim

The invention claimed is:

1. A cell-concentration adjustment device, the device comprising:
an inlet for taking in a cell suspension containing cells at a first concentration;
an outlet for discharging a cell suspension containing cells at a second, desired concentration which is lower than the first concentration at the inlet; and
a flow path which is provided between the inlet and the outlet and is capable of holding a cell suspension,
the flow path comprising
a liquid delivery pump for causing a cell suspension inside to flow;
a cell-concentration measurement instrument for collecting data related to a cell concentration per unit amount of the cell suspension; and
a dilution-liquid container for holding a dilution liquid which is supplied to the flow path to dilute the cell suspension,

wherein the cell-concentration adjustment device further comprises
a controller configured to control at least the liquid delivery pump on the basis of the data obtained by the cell-concentration measurement instrument,
wherein the controller determines, on the basis of the data obtained by the cell-concentration measurement instrument, an amount of the dilution liquid required to bring the cell concentration into the desired concentration, and drives the liquid delivery pump so as to take in the required amount of the dilution liquid into the flow path and mix the cell suspension and the dilution liquid, and
wherein at least a part of the flow path provided between the inlet and the outlet forms a circulation flow path, the liquid delivery pump and the cell-concentration measurement instrument are provided in the circulation flow path, and the controller drives the liquid delivery pump to cause the cell suspension and the dilution liquid to repeatedly flow in the circulation flow path until a variation in the data obtained from the cell-concentration measurement instrument converges to a value within a predetermined range, to thereby mix the cell suspension and the dilution liquid.

2. The cell-concentration adjustment device according to claim 1 , further comprising a buffer tank in the circulation flow path.

3. The cell-concentration adjustment device according to claim 1 , wherein the control unit drives the liquid delivery pump alternately in a forward direction and a reverse direction, to thereby mix the cell suspension and the dilution liquid.

4. The cell-concentration adjustment device according to claim 1 , wherein the cell-concentration measurement instrument measures an intensity of scattered light or transmitted light of light emitted to the cell suspension to collect the data related to the cell concentration as the light intensity, and the controller calculates the cell concentration by comparing the data with a relationship between cell concentration and light intensity which is determined in advance.

5. The cell-concentration adjustment according to claim 1 , wherein the collection of the data related to the cell concentration by the cell-concentration measurement instrument is performed intermittently or continuously in a state where the cell suspension is flowing.

6. The cell-concentration adjustment device according to claim 1 , wherein the controller is configured to control a valve for controlling taking-in of the cell suspension from the inlet and a valve for controlling taking-in of the dilution liquid into the flow path, and the controller controls the liquid delivery pump and the two valves so as to alternately and repeatedly perform the taking-in of the cell suspension and the taking-in of the dilution liquid.

7. An automatic subculture system, comprising:
a first cell culture device for expansion and culture;
a cell-concentration adjustment device; and
a second cell culture device for subculture,
wherein the first cell culture device discharges a cell suspension having a first concentration, the cell-concentration adjustment device dilutes the cell suspension having the first concentration into a uniform cell suspension having a second, desired cell concentration, the second cell culture device inoculates the diluted cell suspension to perform subculture, and
wherein the cell-concentration adjustment device includes an inlet for taking in the cell suspension having the first concentration, an outlet for discharging the cell suspension containing cells at the second, desired concentration which is lower than the first concentration at the inlet, and a flow path which is provided between the inlet and the outlet and is capable of holding a cell suspension,
wherein the flow path being provided with a cell-concentration measurement instrument for collecting data related to a cell concentration per unit amount of a cell suspension, and a dilution-liquid container for holding a dilution liquid which is supplied to the flow path to dilute the cell suspension,
wherein the cell-concentration adjustment device further comprises a controller configured to control a flow of a cell suspension inside the flow path on the basis of the data obtained by the cell-concentration measurement instrument using a liquid delivery pump provided in the first cell culture device or the second cell culture device,
wherein the controller determines, on the basis of the data obtained by the cell-concentration measurement instrument, an amount of the dilution liquid required to bring the cell concentration into the second, desired concentration, and controls the flow of the cell suspension inside the flow path so that the required amount of the dilution liquid is taken in into the flow path and the cell suspension and the dilution liquid are mixed, and
wherein at least a part of the flow path provided between the inlet and the outlet forms a circulation flow path, the liquid delivery pump and the cell-concentration measurement instrument are provided in the circulation flow path, and the controller drives the liquid delivery pump to cause the cell suspension and the dilution liquid to repeatedly flow in the circulation flow path until a variation in the data obtained from the cell-concentration measurement instrument converges to a value within a predetermined range, to thereby mix the cell suspension and the dilution liquid.

8. A method for diluting a cell suspension containing cells at a first concentration into a second, desired concentration, the method comprising:
receiving, via an inlet, a cell suspension containing cells at a first concentration;
intermittently or continuously measuring an intensity of scattered light or transmitted light of light emitted to the cell suspension in a state where the cell suspension is flowing, to thereby collect data related to a cell concentration as the light intensity;
comparing the obtained data with a relationship between cell concentration and light intensity which is determined in advance to convert the data to a cell concentration; and
calculating an amount of a dilution liquid required for dilution to the desired concentration, adding the amount of a dilution liquid to the cell suspension, and mixing the dilution liquid with the cell suspension; and
discharging, via an outlet, a cell suspension containing cells at a second, desired concentration which is lower than the first concentration at the inlet,
wherein said mixing the dilution liquid with the cell suspension further comprises driving a liquid delivery pump to cause the cell suspension and the dilution liquid to repeatedly flow in a circulation flow path until a variation in the measured cell-concentration converges to a value within a predetermined range, to thereby mix the cell suspension and the dilution liquid.

Assignees

• Hitachi High Tech Corp

Inventors

• Shimase Akihiro
• Imai Kazumichi
• Aso Sadamitsu
• Takada Eiichiro
• Kawarai Masako
• Sakurai Toshinari

Classifications

• C12M31/10

  by light emitting elements located inside the reactor, e.g. LED or OLED · CPC title

• C12M41/40 Primary

  of pressure · CPC title

• C12M41/06

  of illumination · CPC title

• C12Q1/06

  Quantitative determination · CPC title

• C12M29/18

  External loop; Means for reintroduction of fermented biomass or liquid percolate (loop type reactors for chemical or physical processes B01J19/2435) · CPC title