US-12618041-B2 - Cell culturing system

US12618041B2US 12618041 B2US12618041 B2US 12618041B2US-12618041-B2

Abstract

A cell culturing system is equipped with a reactor, a flow path through which a culture medium flows into and flows out from the reactor, an optical measurement unit that optically measures cells contained in the culture medium, and a control unit that processes measurement results of the optical measurement unit. The optical measurement unit includes, at different positions in a flow direction of the culture medium, a first measurement unit that performs optical measurements based on of emission a first measurement light, and a second measurement unit that performs optical measurements based on emission of a second measurement light. In addition, a first optical path length of the first measurement light is set to be longer than a second optical path length of the second measurement light.

Inventors

Masatsugu IGARASHI

Assignees

TERUMO KABUSHIKI KAISHA

Dates

Publication Date: 20260505
Application Date: 20230602
Priority Date: 20201224

Claims (14)

1 . A cell culturing system comprising: a reactor configured to culture cells and to allow a culture medium to flow therethrough; an optical measurement unit disposed downstream of the reactor in a flow direction of the culture medium, the optical measurement unit configured to irradiate the culture medium with measurement light and thereby optically measure a state of the culture medium, the optical measurement unit including: a first measurement unit and a second measurement unit arranged at positions in the flow direction of the culture medium, wherein the first measurement unit includes a first optical path section configured to receive the culture medium and a first measurement light transmission, wherein the second measurement unit includes a second optical path section configured to receive the culture medium and a second measurement light transmission, the optical measurement unit; and a processing unit configured to process a measurement result of the optical measurement unit including; when cell culturing is initiated, to calculate number of cells in the reactor using a first measurement result of the first measurement unit; to compare the calculated number of cells to a predetermined switching threshold value; and when the calculated number of cells is greater than or equal to the predetermined switching threshold value, to calculate a revised number of cells in the reactor using a second measurement result of the second measurement unit.
2 . The cell culturing system of claim 1 , wherein the first measurement unit and the second measurement unit arranged at different positions in the flow direction of the culture medium.
3 . The cell culturing system of claim 1 , wherein the processing unit is further configured: to compare the calculated revised number of cells to a predetermined collection value; and to temporarily stop flowing of the culture medium to the reactor and supply to the reactor a stripping solution for stripping off the cells from the reactor.
4 . The cell culturing system of claim 1 , wherein the first measurement unit and the second measurement unit share a light emitting unit that is configured to emit both the first measurement light transmission and the second measurement light transmission.
5 . The cell culturing system of claim 1 , wherein the first measurement light transmission has a first light intensity and the second measurement light transmission as a second light intensity, the second light intensity being greater than the first light intensity.
6 . The cell culturing system of claim 1 , wherein a first optical path length along a first direction of the first measurement light transmission is longer than a second optical path length along a second direction of the second measurement light transmission.
7 . The cell culturing system of claim 1 , wherein the cell culturing system further includes: a first cell arranged in the first optical path section, and a second cell arranged in the second optical path section.
8 . The cell culturing system of claim 7 , wherein the first cell is connected to a tube and the first cell has a first modulus of elasticity that is greater than a second modulus of elasticity of the tube.
9 . The cell culturing system of claim 8 , wherein the first cell includes a first end part configured to connect to the tube, the first end part including an end-side tapered portion having a first end with a first cross-sectional area and configured to couple to the first cell, and a second end with a second cross-sectional area and configured to couple to the tube, the first cross-sectional area being greater than the second cross-sectional area.
10 . The cell culturing system of claim 7 , wherein the second cell is connected to a second tube and the second cell has a first modulus of elasticity that is greater than a second modulus of elasticity of the tube.
11 . The cell culturing system of claim 10 , wherein the second cell includes a second end part configured to connect to the tube, the second end part including an end-side tapered portion having a first end with a first cross-sectional area and configured to couple to the second cell, and a second end with a second cross-sectional area and configured to couple to the tube, the first cross-sectional area being greater than the second cross-sectional area.
12 . The cell culturing system of claim 7 , wherein the first cell and the second cell form a single container, wherein the first cell and the second cell are continuous with each other along the flow direction of the culture medium.
13 . The cell culturing system of claim 12 , wherein the single container includes a flow path transitioning section between the first cell and the second cell, the flow path transitioning section having a first end with a first cross-sectional area, and a second end with a second cross-sectional area, the first cross-sectional area being greater than the second cross-sectional area.
14 . A cell culturing system comprising: a flow path configured to receive culture medium; a reactor in communication with the flow path and configured to culture cells; an optical measurement unit in communication with the flow path downstream of the reactor in a flow direction of the culture medium, the optical measurement unit configured to irradiate the culture medium with measurement light and thereby optically measure a state of the culture medium, the optical measurement unit including: a first measurement unit including a first optical path section configured to receive the culture medium and a first measurement light transmission, and a second measurement unit including a second optical path section configured to receive the culture medium and a second measurement light transmission, a first optical path length along a first direction of the first measurement light transmission being longer than a second optical path length along a second direction of the second measurement light transmission; and a processing unit configured to process a measurement result of the optical measurement unit including; when cell culturing is initiated, to calculate number of cells in the reactor using a first measurement result of the first measurement unit; to compare the calculated number of cells to a predetermined switching threshold value; and when the calculated number of cells is greater than or equal to the predetermined switching threshold value, to calculate a revised number of cells in the reactor using a second measurement result of the second measurement unit.

Description

CROSS-REFERENCED TO RELATED APPLICATIONS The present application is a continuation application of the International Patent Application No. PCT/JP2021/046188 filed on Dec. 15, 2021, which designated the U.S. and claims the benefit of priority from Japanese Patent Application No. JP2020-214579 filed Dec. 24, 2020. The entire disclosures of the above-identified applications are incorporated herein by reference. FIELD The present disclosure relates to a cell culturing system that is used to measure a number of cells flowing from a reactor. BACKGROUND In the practice of regenerative medicine, a treatment may be performed in which biological cells are collected and cultured, and the cultured cells may be administered to a patient. In a cell culturing process for culturing cells, for example, as disclosed in JP 2017-143775 A, a cell culturing system may be used that has a cell culturing container (e.g., reactor)) that includes hollow fibers disposed in an interior of a case. In such a cell culturing system, culturing of cells may be carried out by seeding the cells in the hollow fibers of the reactor, and thereafter, a culture medium may be delivered into the reactor via a flow path connected to the reactor. In this type of cell culturing system, in order to grasp the state of the cell culture and determine the timing at which the cells are collected, it is important to accurately monitor the number of cells that are propagated during culturing. Although it is a general practice to aseptically collect the culture medium inside the cell culturing system through an aseptic filter, the cells are incapable of passing through aseptic filters. For this reason, the culture medium is often collected in an aseptic manner, and the concentration of metabolites of the cells (such as lactic acid, glucose, and oxygen) within the culture medium is measured, and used to indirectly calculate the number of cells that were propagated. Further, as a method of aseptically collecting the cells directly, a circuit tube of the cell culturing system may be sealed over a certain length, and a piece of the tube including the cell-containing culture medium may be collected. Thereafter, the disconnected circuit of the cell culturing system may be connected again using an aseptic joining device. In the instance of indirectly calculating the number of cells as a result of changes in the culturing environment, the cell metabolism can undergo changes and an accurate measurement of the number of cells often cannot be performed. Further, in the case of collecting the tube in which the cell-containing culture medium is included, the operation often becomes complicated, and moreover the culturing operation must be temporarily interrupted, such that cells are lost at the time of sampling. SUMMARY The present disclosure provides a cell culturing system in which it is possible to continuously measure the number of cells in a reactor in a state in which sterility is assured and where a sufficient degree of measurement accuracy can be obtained. A cell culturing system according to the present disclosure includes a reactor configured to culture cells based on a culture medium that flows therethrough, a flow path through which the culture medium flows into and flows out from the reactor, an optical measurement unit disposed in the flow path downstream of the reactor in a flow direction of the culture medium, and a processing unit that may be configured to process a measurement result of the optical measurement unit. The optical measurement unit may be configured to irradiate the culture medium with measurement light and thereby to optically measure a state of the culture medium. The optical measurement unit may include a first measurement unit and a second measurement unit arranged at different positions in the flow direction of the culture medium. The first measurement unit may include a first optical path section through which the culture medium flows together with first measurement light being transmitted therein. The second measurement unit ay include a second optical path section through which the culture medium flows together with second measurement light being transmitted therein. A first optical path length of the first optical path section may be longer than a second optical path length of the second optical path section. The cell culturing system may be configured to continuously measure the number of cells in the reactor in a state in which sterility is assured and where a sufficient degree of measurement accuracy can be obtained. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a perspective view illustrating a structure of an example cell culturing system according to at least one example embodiment of the present disclosure; FIG. 2 is a circuit diagram illustrating an example flow path and an example flow path control mechanism between a culture medium storage unit and a reactor for use with the example cell culturing system illustrated in FIG. 1;