Pressure and circulation culture apparatus and pressure and circulation culture system

• US 9,752,112 B2
• Filed: 02/23/2012
• Issued: 09/05/2017
• Est. Priority Date: 10/09/2009
• Status: Active Grant

• Current Assignee: Purpose Co., Ltd.
• Original Assignee: Purpose Co., Ltd.
• Inventors: Watanabe, Setuo, Murata, Toshimi, Satou, Mituharu, Kinouchi, Ibuki, Takai, Hidetada
• Primary Examiner(s): Beisner, William H
• Assistant Examiner(s): HASSAN, LIBAN M
• Application Number:
  US13/403,186
• Publication Number:
  US 20120156768A1
• Time in Patent Office: 2,021 Days
• Field of Search: 4352891
• CPC Class Codes: C12M 21/08 for producing artificial tissue or for ex-vivo cultivation of tissue prostheses A61F2/00, grafts A61L27/00
• CPC Class Codes: C12M 23/48 Holding appliances; Racks; Supports holding devices for laboratory apparatus B01L9/00
• CPC Class Codes: C12M 27/00 Means for mixing, agitating or circulating fluids in the vessel by introduction of gas C12M29/06, C12M29/14, mixing in general or mixers per se B01F; mixing in apparatus for chemical or physical processes B01J
• CPC Class Codes: C12M 29/14 Pressurized fluid
• CPC Class Codes: C12M 35/04 Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli

Abstract

The pressure and circulation culture apparatus includes a holder (culture chamber part), a pressure transmission part and a culture solution circulation line (culture circuit). The holder reserves culture solution and holds a cultured object. The pressure transmission part communicates with the holder and transmits an external pressure to the culture solution. The culture solution circulation line is connected to the holder via the pressure transmission part, and circulates the culture solution through the holder via the pressure transmission part. Such a structure is included, and an external pressure is made to operate on the culture solution via the pressure transmission part to allow pressure on the cultured object in the holder and circulation of the culture solution in the holder.

Claims

1. A pressure and circulation culture apparatus for culturing a cultured object, and for applying a pressure to a culture solution and releasing the pressure on the culture solution, the pressure and circulation culture apparatus comprising:

an incubator in which a culture environment of the cultured object is maintained;

a pressure generator unit that is outside the incubator; and

a culture unit including a culture unit main body disposed inside the incubator;

wherein the culture unit main body is separable from the culture unit, the culture unit main body culturing the cultured object with the culture unit, and

wherein the culture unit main body comprises;

a culture solution reservoir that reserves the culture solution;

a culture solution circuit that circulates the culture solution from the culture solution reservoir through the culture unit main body;

a holder that holds the cultured object, and the culture solution that circulates through the culture solution circuit;

a pressure transmission part that is in fluid communication with the holder and transmits an external pressure to the culture solution in the holder, wherein the pressure generator unit is connected to the pressure transmission part via a pressure circuit that transmits the external pressure from the pressure generator unit to the pressure transmission part;

a first valve that is disposed at an upstream side of the pressure transmission part, the first valve being in a closed position when a pressure in the pressure transmission part is higher than a predetermined pressure, the closed position of the first valve checking a backflow of the culture solution from the pressure transmission part to the culture solution circuit;

a second valve that is disposed at a downstream side of the holder, the second valve being in a closed position when a pressure in the holder is lower than the predetermined pressure, and the second valve checking a backflow of the culture solution from the culture solution circuit to the holder while the culture solution is drawn into the holder; and

a shut-off valve part that is disposed in the culture solution circuit at a downstream side of the holder and the second valve, the shut-off valve part being switched to a closed position during a process of applying the pressure to the holder, so as to check a circulation of the culture solution, and the shut-off valve part being switched to an open position during a process of circulating the culture solution,

wherein the pressure and circulation culture apparatus further comprises;

a shut-off valve drive unit that attaches to the shut-off valve part, the shut-off valve drive unit opening or closing the shut-off valve part;

a pressure sensor that detects the pressure in the holder, the pressure being applied to the culture solution; and

a processor that is programmed to execute each of the process of applying the pressure and the process of circulating the culture solution, wherein in the process of applying the pressure, the processor is programmed such that the pressure generator unit provides a pressure in the holder that alternates between at least a first target pressure and a second target pressure, the second target pressure being different from the first target pressure, and wherein in the process of circulating the culture solution, the culture solution is made to flow out of the holder after the process of applying the pressure, and then drawn into the holder after flowing out of the holder,

wherein in the process of applying the pressure, the shut-off valve part is closed by the shut-off valve drive unit receiving a control output generated by the processor and the external pressure is applied from the pressure generator unit via the pressure circuit through the pressure transmission part to the holder, so that the pressure in the holder alternates between a first pressurization in which the pressure in the holder reaches and maintains the first target pressure and a second pressurization in which the pressure in the holder reaches and maintains the second target pressure,

wherein after alternating between the first pressurization and the second pressurization, the process of applying the pressure moves to the process of circulating the culture solution,

wherein in the process of circulating the culture solution, the shut-off valve part is released by the shut-off valve drive unit receiving the control output generated by the processor, and wherein in a process of making the culture solution flow out of the holder during the process of circulating the culture solution, the external pressure is applied from the pressure generator unit via the pressure circuit through the pressure transmission part to the culture solution in the holder with the first valve in a closed state and the second valve in an open state for making the culture solution in the holder flow into a downstream side of the culture solution circuit,

wherein in a process of drawing the culture solution into the holder during the process of circulating the culture solution, the external pressure, which is applied from the pressure generator unit via the pressure circuit through the pressure transmission part to the holder, is reduced with the first valve in an open state and the second valve in a closed state, so as to draw the culture solution from a culture solution reservoir side into the holder,

wherein the culture unit main body of the pressure and circulation culture apparatus is separable from the culture unit at the shut-off valve part and the pressure transmission part, and

wherein the cultured object is transportable as a unit with the culture unit main body by making the culture unit main body separate from the culture unit at the shut-off valve part and the pressure transmission part after culture of the cultured object.

2. The pressure and circulation culture apparatus of claim 1,

wherein a pressure transmission space that stores the culture solution is formed in the pressure transmission part,

wherein the pressure transmission part comprises an external pressure operation part which applies the external pressure on a pressure transmission film that is disposed between the pressure transmission space and the external pressure operation part, and

wherein the pressure transmission film is reciprocated by applying and reducing the external pressure, and the external pressure is transmitted to the culture solution.

3. A pressure and circulation culture system for culturing a cultured object, and for applying a pressure to a culture solution and releasing the pressure on the culture solution, the pressure and circulation culture apparatus comprising:

an incubator in which a culture environment of the cultured object is maintained;

a pressure generator unit that is outside the incubator; and

a plurality of culture units disposed inside the incubator, each of the plurality of culture units comprising a culture unit main body;

wherein the culture unit main body is separable from the culture unit, the culture unit main body culturing the cultured object with the culture unit, and

wherein the culture unit main body comprises;

a culture solution reservoir that reserves the culture solution;

a culture solution circuit that circulates the culture solution from the culture solution reservoir through the culture unit main body;

a holder that holds the cultured object, and the culture solution that circulates through the culture solution circuit;

a pressure transmission part that is in fluid communication with the holder and transmits an external pressure to the culture solution in the holder, wherein the pressure generator unit is connected to the pressure transmission part via a pressure circuit that transmits the external pressure from the pressure generator unit to the pressure transmission part;

a first valve that is disposed at an upstream side of the pressure transmission part, the first valve being in a closed position when a pressure in the pressure transmission part is higher than a predetermined pressure, the closed position of the first valve checking a backflow of the culture solution from the transmission part to the culture solution circuit;

a second valve that is disposed at a downstream side of the holder, the second valve being in a closed position when a pressure in the holder is lower than the predetermined pressure, and the second valve checking a backflow of the culture solution from the culture solution circuit to the holder while the culture solution is drawn into the holder; and

a shut-off valve part that is disposed in the culture solution circuit at a downstream side of the holder and the second valve, the shutoff valve part being switched to a closed position during a process of applying the pressure to the holder, so as to check a circulation of the culture solution, and the shut-off valve part being switched to an open position during a process of circulating the culture solution,

wherein the pressure and circulation culture system further comprises;

a shut-off valve drive unit that attaches to the shut-off valve part, the shut-off valve drive unit opening or closing the shut-off valve part;

a pressure sensor that detects the pressure in the holder, the pressure being applied to the culture solution; and

a processor that is programmed to execute each of the process of applying the pressure and the process of circulating the culture solution, wherein in the process of applying the pressure, the processor is programmed such that the pressure generator unit provides a pressure in the holder that alternates between at least a first target pressure and

a second target pressure, the second target pressure being different from the first target pressure, and wherein in the process of circulating the culture solution, the culture solution is made to flow out of the holder after the process of applying the pressure, and then drawn into the holder after flowing out of the holder,

wherein in the process of applying the pressure, the shut-off valve part is closed by the shut-off valve drive unit receiving a control output generated by the processor and the external pressure is applied from the pressure generator unit via the pressure circuit through the pressure transmission part to the holder, so that the pressure in the holder alternates between a first pressurization in which the pressure in the holder reaches and maintains the first target pressure and a second pressurization in which the pressure in the holder reaches and maintains the second target pressure,

wherein after alternating between the first pressurization and the second pressurization, the process of applying the pressure moves to the process of circulating the culture solution,

wherein in the process of circulating the culture solution, the shut-off valve part is released by the shut-off valve drive unit receiving the control output generated by the processor, and wherein in a process of making the culture solution flow out of the holder during the process of circulating the culture solution, the external pressure is applied from the pressure generator unit via the pressure circuit through the pressure transmission part to the culture solution in the holder with the first valve in a closed state and the second valve in an open state for making the culture solution in the holder flow into a downstream side of the culture solution circuit,

wherein in a process of drawing the culture solution into the holder during the process of circulating the culture solution, the external pressure, which is applied from the pressure generator unit via the pressure circuit through the pressure transmission part to the holder, is reduced with the first valve in an open state and the second valve in a closed state, so as to draw the culture solution from a culture solution reservoir side into the holder,

wherein the culture unit main body of the pressure and circulation culture apparatus is separable from the culture unit at the shut-off valve part and the pressure transmission part, and

wherein the cultured object is transportable as a unit with the culture unit main body by making the culture unit main body separate from the culture unit at the shut-off valve part and the pressure transmission part after culture of the culture object.

4. The pressure and circulation culture system of claim 3,

wherein a pressure transmission space that stores the culture solution is formed in the pressure transmission part, wherein the pressure transmission part comprises an external pressure operation part which applies the external pressure on a pressure transmission film that is disposed between the pressure transmission space and the external pressure operation part, and

wherein the pressure transmission film is reciprocated by applying and reducing the external pressure, and the external pressure is transmitted to the culture solution.