• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    A cell culture platform according to the invention comprises culture elements for culturing cells. Each culture element comprises a culture chamber for containing cultured cells and a culture medium in liquid form. The culture element comprises a gas chamber and a gas permeable material between the gas chamber and the culture chamber. The cell culture platform includes a frame structure for mechanically supporting the culture elements and for mechanically supporting a gas supply system capable of autonomously supplying gas without receiving gas from an external source. The gas supply system can be, for example, a gas cartridge. The frame structure includes gas channels for conducting the gas to pass through the gas chambers of the culture elements.
    公开号:FR3045068A3
    申请号:FR1662204
    申请日:2016-12-09
    公开日:2017-06-16
    发明作者:Joose Kreutzer
    申请人:Tty-Saatio；Tampereen yliopisto；
    IPC主号:
    专利说明:

    CELL CULTURE PLATFORM AND CULTURE SYSTEM
    CELLULAR
    Field of the invention
    The present invention relates generally to cell culture. It relates more particularly to a cell culture platform and a cell culture system.
    Context
    A cell culture process can take months and usually involves many laborious manual operations. The culture process may include, for example, changing the culture medium in liquid form every two or three days and / or performing microscopic imaging to monitor the growth of the cultured cells. Whenever operations of a type mentioned above are performed, the cell culture is usually taken out of an incubator. This causes environmental changes within the cell culture, and these environmental changes can cause undesirable effects in the culture process. Living cells are sensitive to the environment where they are grown. Therefore, the aforementioned operations may lead to undesirable cell stimulations, for example due to changes in gas concentrations, temperature and / or light intensity. In addition, mechanical stimulation such as vibration and agitation may cause undesirable effects. It is therefore important to maintain cells under controlled conditions throughout the culture process in order to reliably study cells and tissues.
    For example, research on hypoxia is one of the areas of research that is suffering from environmental changes related to cell cultures. The study of the effect of hypoxic conditions on living cells requires continuous control and management of the gaseous environment of cultured cells. Hypoxia chambers are available commercially but are very expensive to acquire and maintain, and are therefore not available for all research groups. In addition, commercial hypoxia chambers are large hoods that do not provide rapid dynamic changes in the gaseous environment of a cell culture. Stabilizing the gaseous environment after a change may take several hours. In addition, microscopes capable of operating inside hypoxia chambers are also expensive to be used for studies of hypoxia. In the absence of a microscope inside a hypoxia chamber, a cell culture platform must be removed from the hypoxia chamber and, as a corollary, the low oxygen concentration rapidly disappears from the environment. gaseous cell culture. This could create severe cell damage. Therefore, in this area of research, it is very important to maintain cells under the same conditions throughout the culture process. summary
    The following is a simplified summary to provide a basic understanding of certain aspects of various embodiments of the invention. The summary is not a thorough presentation of the invention. It is not intended to identify the key or critical elements of the invention, nor is it intended to delineate the scope of the invention. The following summary merely presents certain concepts of the invention in a simplified form as a prelude to a more detailed description of non-limiting exemplary embodiments of the invention.
    According to the invention, there is provided a new cell culture platform suitable for research on hypoxia, for example. A cell culture platform according to the invention comprises one or more culture elements for culturing cells and a frame structure for mechanically supporting the culture elements. Each of the culture elements comprises: a culture chamber for containing cultured cells and a culture medium in liquid form, a gas chamber for containing gas, and a gas permeable material between the gas chamber and the chamber. of culture.
    The frame structure is configured to mechanically support not only the growing elements but also a gas supply system which is capable of supplying gas having a desired composition without receiving the gas from an external source. The frame structure includes gas channels for receiving gas from the gas supply system and conducting the gas to pass through the gas chambers of the culture elements. The gas supply system may be, for example but not necessarily, a gas cartridge or other replaceable and / or refillable container for the pressurized gas.
    According to the invention, a new cell culture system is also provided. A cell culture system according to the invention comprises: a cell culture platform according to the invention, and a gas supply system mechanically supported by the frame structure of the cell culture platform.
    The gas supply system may include a replaceable container for the pressurized gas.
    The gas supply system may include a refillable container for pressurized gas, the refillable container including a fill valve.
    The cell culture system can be portable, so that it can be placed, for example, on a sample table of a microscope so that the gas supply system operates during microscopic imaging. Thus, microscopic imaging that may be required to monitor cell growth does not cause change in the gaseous environment of the cells.
    The volume of the aforementioned gas chamber is advantageously small in order to be able to make rapid changes, if necessary, in the gaseous environment of the cells. For example, the volume of the gas chamber may be from about 0.5 to about 3 times the volume of the culture chamber for containing the cultured cells.
    A number of examples of non-limiting embodiments of the invention may include the following additional features considered singly or in combination: the frame structure comprises a well plate comprising wells which are each capable of containing one of the culture elements, and a lid including the gas channels and for closing the wells when placed above the well plate; the well plate comprises a cavity intended to contain the gas supply system; the frame structure is configured to mechanically support another gas supply system, and the gas channels comprise a selection valve system for selecting which of the gas supply systems is allowed to deliver the gas to the chambers. gas of the elements of culture; each of the culture elements comprises: a first reservoir intended to contain the culture medium in liquid form, a first liquid conduit intended to conduct the culture medium in liquid form from the first reservoir to the culture chamber, and a second liquid conduit for conducting the culture medium in liquid form out of the culture chamber of the culture element into consideration; each of the culture elements comprises a second reservoir connected to the second liquid conduit of the culture element in consideration; each of the culture elements comprises: a first gas conduit for conducting gas from the gas channels to the gas chamber of the culture element in consideration, and a second gas conduit for conducting the gas out of the chamber to gas of the element of culture in consideration; the gas chamber of each of the culture elements surrounds the culture chamber of the culture element in consideration, and a wall between the gas chamber and the culture chamber comprises the gas-permeable material; each of the culture elements includes portions stacked one above the other such that each of the portions has a constant cross-sectional profile when a sectional plane is perpendicular to a direction in which the portions are each other; above others; - the frame structure constitutes a background for each of the elements of culture; - The frame structure and the culture elements are at least partially made of one or more transparent materials so as to allow optical inspection of the cultured cells; the transparent material (s) comprises / comprise one or more of the following elements: polystyrene, polystyrene with copolymers, polyvinyl chloride, polyvinyl chloride with copolymers, polyethylene, polystyrene-acrylonitrile, polypropylene, polyvinylidene chloride silicone elastomer; the cell culture platform further comprises an electrically operated heating element for controlling the temperature of the cultured cells; the gas-permeable material comprises polydimethylsiloxane; - The cell culture platform further comprises a controllable valve for controlling a flow of gas through the gas chambers of the culture elements; the cell culture platform further comprises a reflux barrier for preventing ambient air from flowing into the gas chambers of the culture elements in a direction opposite to a flow direction of the gas supplied by the control system. gas supply; a volume of the gas chamber of each of the culture elements is from 0.5 to 3 times, preferably from 0.5 to 3 times, a volume of the culture chamber of the culture element in consideration;
    Various examples of non-limiting embodiments of the invention both in terms of constructions and methods of operation, in conjunction with other objects and advantages thereof, will be better understood from the following description of examples of specific embodiments discussed in connection with the accompanying drawings.
    The verbs "understand" and "include" are used in this document as open limits that do not exclude or require the existence of equally undescribed features. The features set out above may be combined with each other and freely unless explicitly stated otherwise.
    Brief description of the figures
    Examples of non-limiting embodiments of the invention and their advantages are explained in more detail hereinafter with reference to the drawings, in which: FIG. 1a illustrates the mechanical structure of a cell culture system according to an example non-limiting embodiment of the invention, Figure lb illustrates the operation of a cell culture system according to an exemplary non-limiting embodiment of the invention, and Figures 2a-2g illustrate the mechanical structure of the elements. of culture of the cell culture system shown in Figure la.
    Description of Examples of Non-Limiting Embodiments
    The specific examples provided in the following description are not to be construed as limiting the scope and / or applicability of the appended claims. The lists and groups of examples provided in the description below are not exhaustive unless explicitly stated otherwise.
    Figure la shows an isometric view of a cell culture system according to an exemplary non-limiting embodiment of the invention. The cell culture system comprises a cell culture platform 101 and a gas supply system 111 which is capable of providing gas having a desired composition without receiving gas from an external source. The cell culture platform 101 comprises culture elements for culturing cells. In Figure la, one of the culture elements is indicated by a numeral mark 102. The other culture elements are not shown in Figure la. The cell culture platform 101 further comprises a frame structure 103 for mechanically supporting the culture elements and the gas supply system 111. The frame structure 103 comprises gas channels 104 for receiving the gas. of the gas supply system 111 and to conduct the gas to pass through the culture elements. In this example case, the culture elements, such as the culture element 102, are separate components with respect to the frame structure 103. It is also possible, however, that each culture element is an integral part of the structure of the frame. frame.
    The frame structure 103 is advantageously at least partially made of one or more transparent materials. Correspondingly, each culture element is advantageously at least partially made of one or more transparent materials. In this example case, cultured cells can be imaged and / or examined using optical microscopy techniques. The transparent material (s) may include, for example, polystyrene, polystyrene with copolymers, polyvinyl chloride, polyvinyl chloride with copolymers, polyethylene, polystyrene-acrylonitrile, polypropylene polyvinylidene chloride, polycarbonate, cyclic olefin copolymer and / or silicone elastomer.
    In the exemplary cell culture platform 101 shown in FIG. 1a, the frame structure 103 comprises a well plate 105 and a lid 106. The well plate 105 includes wells that are each capable of containing the well. one of the culture elements, and the lid 106 comprises the aforementioned gas channels 104. The lid 106 closes the wells when the lid is placed above the well plate, as shown in FIG. The physical dimensions of the well plate 105 are advantageously similar to the physical dimensions of a standard laboratory well plate so that the cell culture system can be easily placed, for example, on a sample table of a microscope. and / or on another instrument. The length L of the cell culture platform 101 may be, for example, in the range of 86 mm to 128 mm, the width W of the cell culture platform 101 may be, for example, in the range of 60 mm to 86 mm, and the height H of the cell culture platform 101 may be, for example, in the range of 18 mm to 45 mm.
    In the exemplary cell culture system of Fig. La, the gas supply system 111 comprises a replaceable container 114 which contains pressurized gas. The replaceable container 114 may be, for example, a gas cartridge. The replaceable container 114 may contain, for example, 12 g of gas having a volume of about 7 liters at normal temperature and pressure "TPN". With proper design of the cell culture system, the aforementioned volume may be sufficient to maintain sufficient gas flow through individual culture elements for several days. The culture elements are advantageously designed so that a low gas flow, for example, of about 100 μΐ / min, is sufficient to maintain the desired gaseous environment within the culture elements. With the use of the replaceable container 114, there is no need to use external hoses and / or any wires connected to an external system. As a result, the cell culture system is portable and the gaseous environment can still be maintained because the gas supply system 111 is capable of operating autonomously without connections to an external system. Thus, for example, microscopic imaging that may be required to monitor the growth of cultured cells does not cause a change in the gaseous environment of the cells. In the case example illustrated in FIG. 1a, the frame structure 103 includes a cavity for the replaceable container 114. It is also sometimes possible for the frame structure 103 to include means for attaching a pressurized gas container to a container. outer surface of the frame structure.
    It should be noted that the aforementioned replaceable container 114 is not the only possible choice for the gas supply system 111. It is also possible that the gas supply system 111 includes a refillable container for the pressurized gas . The refillable vessel includes a fill valve for filling from an external source such as a gas cylinder and / or a gas mixer. Furthermore, it is also possible that the gas supply system of a cell culture system according to an exemplary embodiment of the invention comprises a chamber for containing liquids which interact so that the desired gas is generated.
    The flow of gas in a cell culture system according to an exemplary non-limiting embodiment of the invention is illustrated in FIG. The cell culture platform of the cell culture system comprises the gas channels 104 for receiving the gas from the gas supply system 111 and driving the gas to pass through the culture elements of which the One is indicated by numeral 102. The cell culture platform may include a controllable valve 109 for controlling the flow of gas through the culture elements. In the exemplary cell culture system shown in FIG. 1a, the controllable valve may be in an element 115. The cell culture platform may further include a reflux barrier to prevent ambient air from passing through the cell culture system. the culture elements in a direction opposite to the flow direction of the gas supplied by the gas supply system. In Figs 1a and 1b, the reflux barrier is indicated by a numeral 110.
    In a cell culture platform according to an exemplary non-limiting embodiment of the invention, the frame structure is configured to mechanically support two or more autonomous gas supply systems. In this example case, the gas channels comprise a selection valve system for selecting which of the gas supply systems is allowed to deliver gas to the culture elements. In Fig. 1b, the selection valve system is indicated by a numeral 107. By using, for example, two replaceable containers, for example, gas cartridges, supported by the frame structure, it is possible to maintain a desired gaseous environment of cultured cells during the change of a replaceable container. In addition, one of the gas supply systems may contain a gas different from another of the gas supply systems. In this case example, it is possible to modify the gaseous environment of the cells in a controlled manner. For example, in conjunction with hypoxia research, it is possible to perform reoxygenation experiments, so that two different gas concentrations are relayed.
    In a cell culture platform according to an exemplary non-limiting embodiment, the gas channels comprise an interface 113 for connection to an external gas supply. In this example case, an external gas cylinder and / or a gas mixer can be connected to the cell culture platform to avoid consumption of gas from a replaceable container and / or or rechargeable, for example, a gas cartridge. In the exemplary cell culture system shown in FIG. 1a, an external gas supply connection interface may be in element 115.
    A cell culture platform according to an exemplary non-limiting embodiment includes an electrically operated heating element for controlling the temperature of the cultured cells. In the exemplary cell culture system shown in FIG. 1a, the heating element is indicated with a numeral 108. The heating element 108 may comprise, for example, resistance wires which are so thin that they not interfere, for example, with microscope imaging and other optical operations. The heating element 108 may, for example, operate by means of a battery, and the frame structure 103 may be arranged to mechanically support the battery.
    Figure 2a illustrates the culture element 102 shown in Figure la. Figures 2b, 2c and 2d show a section obtained by cutting the culture element 102 along an axis A-A shown in Figure 2a. The sectional plane relating to FIGS. 2b-2d is parallel to the xz plane of a coordinate system 199. In FIGS. 2b and 2d, the section surfaces are indicated by oblique hatching. In Figure 2b, the inner shapes are presented in the same manner as in Figure 2a. Figures 2e, 2f and 2g show a section obtained by cutting the culture element 102 along an axis B-B shown in Figure 2a. The sectional plane relating to FIGS. 2e-2g is parallel to the plane yz of the coordinate system 199. The viewing directions relating to FIGS. 1a and 2a-2g are illustrated by the coordinate system 199. The culture element 102 comprises a culture chamber for containing the cultured cells and a culture medium in liquid form. The growth chamber is indicated by a numeral reference 220 in FIGS. 2a, 2d, 2f and 2g. In Fig. 2a, the cultured cells are illustrated by a grid area 255. The culture element 102 includes a gas chamber for containing the gas that is used to provide the desired gaseous environment for the cultured cells. The gas chamber is indicated by a numeral 221 in Figures 2a, 2c, 2f and 2g. The culture element 102 comprises a gas permeable material between the gas chamber 221 and the culture chamber 220. The gas permeable material is indicated by a numeral 222 in Figures 2a, 2e and 2f. The gas-permeable material may comprise, for example, polydimethylsiloxane. The exemplary culture element 102 further comprises a first reservoir for containing the culture medium in liquid form, a first liquid conduit for conducting the culture medium in liquid form from the first reservoir to the culture chamber 220 a second liquid conduit for conducting the culture medium in liquid form out of the culture chamber 220, and a second reservoir 226 connected to the second liquid conduit. The first aforementioned tank is indicated by a numeral 223 on FIGS. 2a-2f, the first aforementioned liquid conduit is indicated by a numeral 224 in FIGS. 2b and 2d, the aforementioned second liquid conduit is indicated by a numerical marker 225 in Figures 2b and 2d, and the second aforementioned tank is indicated by a numeral 226 in Figures 2a-2d. In Fig. 2d, the culture medium in liquid form is indicated by a numeral numeral 250 and the flow of the culture medium in liquid form from the first reservoir 223 through the culture chamber 220 to the second reservoir 226 is illustrated by a curved linear arrow. In the exemplary situation shown in FIG. 2d, the flow is based on a hydrostatic pressure difference between the first and second tanks 223 and 226. As shown in FIG. 2d, the frame structure 103 constitutes the bottom of the However, it is also possible for the cell culture element 102 to have its own bottom element. The exemplary culture element 102 further includes a first gas conduit for conducting the gas from the gas channels 104 shown in Figs. 1a and 1b to the gas chamber 221 and a second gas conduit for conducting the gas. gas out of the gas chamber 221. The first gas pipe is indicated by a numeral 227 in Figures 2a, 2b, 2c, 2e, 2f and 2g, and the second gas pipe is indicated by a numeral 228 on Figures 2a, 2e, 2f and 2g. In Figures 2f and 2g, the gas flow is illustrated by curved linear arrows.
    As shown in FIG. 2a, the gas chamber 221 of the culture element 102 surrounds the culture chamber 220, and a wall between the gas chamber and the culture chamber comprises the gas permeable material 222. As shown in FIG. Figure 2a, the gas chamber has an elongated shape such that the gas chamber is actually a gas channel and thus, the volume of the gas chamber 221 may be relatively small. The small volume of the gas chamber facilitates rapid changes, if necessary, in the gaseous environment of the cultured cells. For example, the volume of the gas chamber 221 may be from about 0.5 to about 3 times the volume of the culture chamber 220 for containing the cells.
    The mechanical structure of the exemplary cultivation element 102 shown in Figures 2a-2g includes portions stacked one above the other. In FIGS. 2a-2g, the abovementioned parts are indicated by the numerical references 229, 230 and 231. As shown in FIGS. 2a-2g, the parts 229-231 are stacked one above the other in the z direction of the As shown in FIGS. 2a-2g, each of the portions 229-231 has a constant cross-sectional profile when a cutting plane is perpendicular to the z-axis of the coordinate system 199.
    The specific non-limiting examples provided in the description above should not be construed as limiting the scope and / or applicability of the appended claims. The lists and groups of examples provided in the description above are not exhaustive unless explicitly stated otherwise.
    权利要求:
    Claims (19)
    [1]
    Claims:
    A cell culture platform (101) comprising one or more culture elements (102) for culturing the cells, each of the culture elements comprising: a culture chamber (220) for containing cultured cells and a culture medium; culture in liquid form, a gas chamber (221) for containing gas, and a gas permeable material (222) between the gas chamber and the culture chamber, characterized in that the cell culture platform comprises further, a frame structure (103) for mechanically supporting the culture elements and mechanically supporting a gas supply system capable of supplying the gas without receiving the gas from an external source, and the frame structure comprises gas channels (104) for receiving gas from the gas supply system and for conducting the gas to pass through the gas chambers of the culture elements.
    [2]
    The cell culture platform of claim 1, wherein the frame structure comprises a well plate (105) comprising wells which are each capable of containing one of the culture elements, and a lid (106). comprising the gas channels and for closing the wells when placed above the well plate.
    [3]
    The cell culture platform of claim 2, wherein the well plate (105) comprises a cavity for containing the gas supply system.
    [4]
    The cell culture platform of any one of claims 1 to 3, wherein the frame structure is configured to mechanically support another gas supply system, and the gas channels comprise a gas valve system. selection (107) for selecting which of the gas supply systems is allowed to deliver the gas to the gas chambers of the culture elements.
    [5]
    The cell culture platform according to any one of claims 1 to 3, wherein each of the culture elements (102) comprises: a first reservoir (223) for containing the culture medium in liquid form, a first liquid conduit (224) for conducting the culture medium in liquid form from the first reservoir to the culture chamber in consideration, and a second liquid conduit (225) for conducting the culture medium in liquid form out of the chamber of culture of the element of culture in consideration.
    [6]
    The cell culture platform of claim 5, wherein each of the culture elements comprises a second reservoir (226) connected to the second liquid conduit of the culture element in consideration.
    [7]
    The cell culture platform according to any one of claims 1 to 3, wherein each of the culture elements comprises: a first gas conduit (227) for conducting gas from the gas channels to the gas chamber ( 221) of the culture element in consideration, and a second gas conduit (228) for conducting the gas out of the gas chamber of the culture element into consideration.
    [8]
    A cell culture platform according to any one of claims 1 to 3, wherein the gas chamber (221) of each of the culture elements surrounds the culture chamber (220) of the culture element into consideration. and a wall between the gas chamber and the culture chamber comprises the gas permeable material.
    [9]
    A cell culture platform according to any one of claims 1 to 3, wherein each of the culture elements comprises portions (229-231) stacked one above the other so that each of the parts has a constant cross-sectional profile when a sectional plane is perpendicular to a direction in which the parts are above each other.
    [10]
    A cell culture platform according to any one of claims 1 to 3, wherein the frame structure constitutes a background for each of the culture elements.
    [11]
    The cell culture platform according to any one of claims 1 to 3, wherein the frame structure and the culture elements are at least partially made of one or more transparent materials so as to enable the optical inspection. cultured cells. 1 polyethylene, polystyrene-acrylonitrile, polypropylene, polyvinylidene chloride, silicone elastomer.
    [13]
    The cell culture platform of any one of claims 1 to 3, wherein the cell culture platform further comprises an electrically operated heating element (108) for controlling the temperature of the cultured cells.
    [14]
    The cell culture platform of any one of claims 1 to 3, wherein the gas permeable material comprises polydimethylsiloxane.
    [15]
    The cell culture platform of any one of claims 1 to 3, wherein the cell culture platform further comprises a controllable valve (109) for controlling a flow of gas through the gas chambers of the elements of culture.
    [16]
    The cell culture platform of any one of claims 1 to 3, wherein the cell culture platform further comprises a reflux barrier (110) for preventing ambient air from flowing into the cells. gas chambers of the culture elements in a direction opposite to a flow direction of the gas supplied by the gas supply system.
    [17]
    The cell culture platform according to any one of claims 1 to 3, wherein a volume of the gas chamber of each of the culture elements is 0.5 to 3 times, preferably 0.5 to 3 times, a volume of the culture chamber of the culture element in consideration.
    [18]
    A cell culture system comprising: a cell culture platform (101) according to any one of claims 1 to 3, and a gas supply system (111) mechanically supported by the frame structure of the platform of cell culture.
    [19]
    The cell culture system of claim 18, wherein the gas supply system comprises a replaceable container (114) for the pressurized gas.
    [20]
    The cell culture system of claim 18, wherein the gas supply system comprises a refillable container for pressurized gas, the refillable container comprising a fill valve. A cell culture platform according to claim 11, wherein the transparent material (s) comprises / comprises one or more of the following: polystyrene, polystyrene with copolymers, polyvinyl chloride, polyvinyl chloride with copolymers,
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    引用文献:
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    法律状态:
    2017-12-18| PLFP| Fee payment|Year of fee payment: 2 |
    2018-12-14| PLFP| Fee payment|Year of fee payment: 3 |
    2019-10-25| TP| Transmission of property|Owner name: BIOGENIUM MICROSYSTEMS OY, FI Effective date: 20190917 |
    2019-12-16| PLFP| Fee payment|Year of fee payment: 4 |
    2020-12-10| PLFP| Fee payment|Year of fee payment: 5 |
    2021-12-15| PLFP| Fee payment|Year of fee payment: 6 |
    优先权:
    申请号 | 申请日 | 专利标题
    FI20155930|2015-12-09|
    FI20155930|2015-12-09|
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