Cell membrane sheet culture apparatus and preparation method

Abstract

This disclosure provides a cell sheet culture apparatus and preparation method, relating to the field of cell culture technology. The cell sheet culture apparatus includes a semiconductor cooling device and one or more culture containers configured to culture cell sheets. The semiconductor cooling device includes a first insulating substrate, a second insulating substrate, and at least one semiconductor thermocouple disposed between the first and second insulating substrates. The one or more culture containers are disposed on the side of the first insulating substrate away from the second insulating substrate.

Description

Technical Field

[0001] This disclosure relates to the field of cell culture technology, and in particular to a cell membrane culture apparatus and preparation method. Background Technology

[0002] Cell membranes are membrane-like tissues formed by the connection between the extracellular matrix secreted by cells and the cells themselves. Cell membranes have high cell density, uniform cell distribution, and homogeneous texture, and can be used for the repair of organs or tissues such as skin, cartilage, cornea, and myocardium. Summary of the Invention

[0003] In related technologies, cell sheets can be prepared using temperature-sensitive culture dishes coated with a temperature-sensitive polymer layer. The temperature of the temperature-sensitive culture dish needs to be adjusted during the preparation of cell sheets.

[0004] In view of the above, the present disclosure provides the following technical solutions.

[0005] According to one aspect of this disclosure, a cell membrane culture apparatus is provided, comprising: a semiconductor cooling device, and one or more culture containers configured to culture cell membranes. The semiconductor cooling device includes a first insulating substrate, a second insulating substrate, and at least one semiconductor thermocouple disposed between the first insulating substrate and the second insulating substrate. The one or more culture containers are disposed on the side of the first insulating substrate away from the second insulating substrate.

[0006] In some embodiments, the semiconductor cooling device further includes: a heat-conducting component disposed on the side of the first insulating substrate away from the second insulating substrate, wherein the one or more culture containers are disposed on the heat-conducting component.

[0007] In some embodiments, the heat-conducting component has one or more grooves, one of which contains a culture container.

[0008] In some embodiments, the projection of the first insulating substrate onto the second insulating substrate is within the projection of the thermally conductive component onto the second insulating substrate.

[0009] In some embodiments, at least one of the culture containers has a temperature-sensitive polymer layer disposed on the bottom side away from the first insulating substrate, and the surface energy of the temperature-sensitive polymer layer changes monotonically with temperature changes.

[0010] In some embodiments, the surface energy of the temperature-sensitive polymer layer increases with increasing temperature.

[0011] In some embodiments, the at least one semiconductor thermocouple includes N semiconductor thermocouples, where N is an integer greater than or equal to 2, wherein: each semiconductor thermocouple includes a first semiconductor portion and a second semiconductor portion with different conductivity types, each of the first semiconductor portion and the second semiconductor portion includes a first surface and a second surface disposed opposite to each other, the first surface being closer to the first insulating substrate than the second surface, the first surface of the first semiconductor portion and the first surface of the second semiconductor portion being connected by a first metal member, and the second surface of the first semiconductor portion of the i-th semiconductor thermocouple and the second surface of the second semiconductor portion of the (i+1)-th semiconductor thermocouple being connected by a second metal member, wherein 1≤i≤N-1.

[0012] In some embodiments, the semiconductor cooling device further includes a first switch, a first power supply, and a second power supply, wherein: a first terminal of the first power supply is connected to a second surface of a second semiconductor portion of a first semiconductor thermocouple via a third metal member; the second terminal of the first power supply is configured to be connected to a second surface of a first semiconductor portion of an Nth semiconductor thermocouple via the first switch and a fourth metal member; a second terminal of the second power supply is connected to a second surface of a second semiconductor portion of a first semiconductor thermocouple via the third metal member; a first terminal of the second power supply is configured to be connected to a second surface of a first semiconductor portion of an Nth semiconductor thermocouple via the first switch and the fourth metal member; the first switch is configured to connect to one of the second terminal of the first power supply and the first terminal of the second power supply in response to a user operation; wherein one of the first terminal and the second terminal is a positive terminal and the other is a negative terminal.

[0013] In some embodiments, the semiconductor cooling device further includes: a temperature monitoring component configured to monitor the temperature of the first insulating substrate; and a controller configured to control the state of the first switch based on the temperature of the first insulating substrate.

[0014] In some embodiments, the controller is configured to: control the first switch to close when the temperature of the first insulating substrate is within a preset temperature range; control the first switch to open when the temperature of the first insulating substrate is not within the preset temperature range; or control the first switch to switch from a connection to one of the second pole of the first power supply and the first pole of the second power supply to a connection to the other of the second pole of the first power supply and the first pole of the second power supply.

[0015] In some embodiments, the semiconductor cooling device further includes: a second switch connected between the first power supply and N semiconductor thermocouples, and a second power supply connected between the second power supply and N semiconductor thermocouples; the controller is configured to: control the second switch to close when the temperature of the first insulating substrate is within a preset temperature range; and control at least one of the first switch and the second switch to open when the temperature of the first insulating substrate is not within the preset temperature range.

[0016] In some embodiments, the semiconductor cooling device further includes: a voltage divider adjustable element connected between the first power supply and N semiconductor thermocouples, and connected between the second power supply and N semiconductor thermocouples.

[0017] In some embodiments, the semiconductor cooling device further includes a heat dissipation component disposed on the side of the second insulating substrate away from the first insulating substrate.

[0018] In some embodiments, the semiconductor cooler is disposed separately from the one or more culture vessels.

[0019] According to another aspect of the present disclosure, a method for preparing a cell sheet based on the cell sheet culture apparatus described in any of the above embodiments is provided. The method for preparing the cell sheet includes: adjusting the temperature of a first insulating substrate of a semiconductor refrigeration device to a first temperature so that a plurality of cells in the culture container form a cell sheet adsorbed onto a temperature-sensitive polymer layer; and adjusting the temperature of the first insulating substrate to a second temperature different from the first temperature so that the cell sheet separates from the temperature-sensitive polymer layer.

[0020] In some embodiments, the second temperature is lower than the first temperature.

[0021] The cell sheet culture apparatus provided in this disclosure includes a semiconductor cooling device. Using a semiconductor cooling device to regulate the temperature of the culture container reduces the influence of ambient temperature on the culture container's temperature, resulting in more accurate temperature regulation. Furthermore, the semiconductor cooling device provides rapid cooling or heating, enabling faster temperature control. Additionally, the semiconductor cooling device is easily miniaturized, allowing the cell sheet culture apparatus to be used in space-constrained environments such as clean benches or biosafety cabinets where cell manipulation is required.

[0022] Other features, aspects, and advantages of this disclosure will become clear from the following detailed description of exemplary embodiments with reference to the accompanying drawings. Attached Figure Description

[0023] The accompanying drawings form part of this specification, depicting exemplary embodiments of the present disclosure and, together with the specification, serving to explain the principles of the present disclosure. In the drawings:

[0024] Figure 1 This is a schematic diagram illustrating the structure of a cell membrane culture apparatus according to an embodiment of the present disclosure;

[0025] Figure 2 This is a schematic diagram illustrating the structure of a semiconductor cooling device according to one implementation of the present disclosure;

[0026] Figure 3 This is a schematic diagram illustrating the structure of a semiconductor cooling device according to another implementation of the present disclosure;

[0027] Figure 4 This is a schematic diagram of the structure of a semiconductor cooling device according to yet another implementation of the present disclosure;

[0028] Figure 5 This is a schematic diagram illustrating the structure of a semiconductor cooling device according to another implementation of the present disclosure;

[0029] Figure 6 This is a schematic flowchart illustrating a method for preparing a cell membrane sheet according to an embodiment of the present disclosure.

[0030] It should be understood that the dimensions of the various parts shown in the accompanying drawings are not necessarily drawn to actual scale. Furthermore, the same or similar reference numerals denote the same or similar components. Detailed Implementation

[0031] Various exemplary embodiments of the present disclosure will now be described in detail with reference to the accompanying drawings. The descriptions of the exemplary embodiments are merely illustrative and are in no way intended to limit the present disclosure or its application or use. The present disclosure may be implemented in many different forms and is not limited to the embodiments described herein. These embodiments are provided so that the present disclosure will be thorough and complete, and will fully express the scope of the disclosure to those skilled in the art. It should be noted that, unless specifically stated otherwise, the relative arrangement of components and steps, the composition of materials, numerical expressions, and values ​​set forth in these embodiments should be interpreted as exemplary only and not as limiting.

[0032] The terms "first," "second," and similar words used in this disclosure do not indicate any order, quantity, or importance, but are merely used to distinguish different parts. Words such as "including" or "containing" mean that the element preceding the word encompasses the element listed after the word, and do not exclude the possibility of encompassing other elements as well. Terms such as "above" and "below" are used only to indicate relative positional relationships, and these relative positional relationships may also change accordingly when the absolute position of the described object changes.

[0033] In this disclosure, when a specific component is described as being located between a first component and a second component, an intermediary component may or may not be present between the specific component and the first or second component. When a specific component is described as connecting to other components, the specific component may be directly connected to the other components without having an intermediary component, or it may not be directly connected to the other components but may have an intermediary component.

[0034] All terms used in this disclosure (including technical or scientific terms) have the same meaning as understood by one of ordinary skill in the art to which this disclosure pertains, unless otherwise specifically defined. It should also be understood that terms defined in a general dictionary, such as a dictionary, should be interpreted as having a meaning consistent with their meaning in the context of the relevant art, and not as having an idealized or highly formalized meaning, unless expressly defined herein.

[0035] Techniques, methods, and equipment known to those skilled in the art may not be discussed in detail, but where appropriate, such techniques, methods, and equipment should be considered part of the specification.

[0036] This disclosure proposes combining semiconductor refrigeration technology with the preparation of cell membrane sheets, which will be described below with reference to different embodiments.

[0037] Figure 1 This is a schematic diagram illustrating the structure of a cell membrane culture apparatus according to an embodiment of the present disclosure.

[0038] like Figure 1 As shown, the cell sheet culture apparatus may include a semiconductor cooling device 10 and one or more culture containers 30. Here, Figure 1 Only one culture container 30 is shown schematically.

[0039] The culture vessel 30 is configured to culture cell membrane sheets. The semiconductor cooling device 10 is configured to regulate the temperature of the culture vessel 30.

[0040] The semiconductor cooling device 10 includes a first insulating substrate 11, a second insulating substrate 12, and at least one semiconductor thermocouple 13. The semiconductor thermocouple 13 is disposed between the first insulating substrate 11 and the second insulating substrate 12. In some embodiments, the semiconductor cooling device 10 may include N semiconductor thermocouples 13, where N is an integer greater than or equal to 2. Figure 1 Three semiconductor thermocouples 13 are schematically shown in the figure.

[0041] See Figure 1Each semiconductor thermocouple 13 may include a first semiconductor portion 131 and a second semiconductor portion 132 with different conductivity types. An insulating material may be filled between the first semiconductor portion 131 and the second semiconductor portion 132. For example, the first semiconductor portion 131 may be an N-type semiconductor, while the second semiconductor portion 132 may be a P-type semiconductor. Alternatively, the first semiconductor portion 131 may be a P-type semiconductor, while the second semiconductor portion 132 may be an N-type semiconductor.

[0042] Each of the first semiconductor section 131 and the second semiconductor section 132 includes a first surface S1 and a second surface S2 disposed opposite to each other. Here, the first surface S1 is closer to the first insulating substrate 11 than the second surface S2. Taking the first semiconductor section 131 as an example, see... Figure 1 The first surface S1 of the first semiconductor section 131 can be, for example, the upper surface of the first semiconductor section 131, and the second surface S2 of the first semiconductor section 131 can be, for example, the lower surface of the first semiconductor section 131.

[0043] The first surface S1 of the first semiconductor unit 131 and the first surface S1 of the second semiconductor 132 can be connected by a first metal member M1. The material of the first metal member M1 may include copper, iron, aluminum, or an alloy of the aforementioned materials.

[0044] The second surface S2 of the first semiconductor portion 131 of the i-th semiconductor thermocouple 13 and the second surface S2 of the second semiconductor portion 132 of the (i+1)-th semiconductor thermocouple 13 are connected by a second metal member M2. Here, 1≤i≤N-1. The material of the second metal member M2 may include copper, iron, aluminum, or an alloy of the aforementioned materials. For example, the first semiconductor thermocouple 13 may be the leftmost semiconductor thermocouple 13, the Nth semiconductor thermocouple 13 may be the rightmost semiconductor thermocouple 13, and vice versa.

[0045] When the semiconductor thermocouple 13 is connected in the circuit, according to the Peltier effect, one of the first insulating substrate 11 and the second insulating substrate 12 is the cold end and the other is the hot end. In some embodiments, the materials of the first insulating substrate 11 and the second insulating substrate 12 may include, but are not limited to, ceramic materials with good thermal conductivity and electrical insulation.

[0046] The culture container 30 is disposed on the side of the first insulating substrate 11 away from the second insulating substrate 12. In some embodiments, the culture container 30 may include, but is not limited to, a culture dish, a culture flask, or a culture plate. As some implementations, the shape of the culture container 30 may include, but is not limited to, a circle or a square.

[0047] In some implementations, the thermoelectric cooler 10 and the culture vessel 30 can be fixed together. For example, the culture vessel 30 can be fixed together with the first insulating substrate 11. In other implementations, the thermoelectric cooler and the culture vessel 30 can be separate, that is, they are not fixed together.

[0048] In some embodiments, a temperature-sensitive polymer layer 21 is disposed on the bottom side of at least one culture container 30 away from the first insulating substrate 11. Here, the surface energy of the temperature-sensitive polymer layer 21 changes monotonically with temperature, for example, monotonically increasing or monotonically decreasing with increasing temperature.

[0049] In some implementations, the surface energy of the thermosensitive polymer layer 21 can increase with increasing temperature. For example, at higher temperatures suitable for cell culture (e.g., around 37°C), the thermosensitive polymer exhibits a coiled shape. In this case, the high surface energy of the thermosensitive polymer results in hydrophobicity, which is beneficial for cell adhesion. Conversely, at lower temperatures (e.g., approximately 15°C to approximately 25°C), the molecular chain structure of the thermosensitive polymer exhibits an extended shape. In this case, the low surface energy of the thermosensitive polymer results in hydrophilicity, which is detrimental to cell adhesion. As an example, the material of the thermosensitive polymer layer 21 may include one or more of the following materials: poly(N-isopropylacrylamide) or poly(N-n-propylacrylamide).

[0050] In some other implementations, the surface energy of the temperature-sensitive polymer layer 21 can decrease as the temperature increases.

[0051] When preparing cell sheets using the aforementioned cell sheet culture apparatus, the temperature of the first insulating substrate of the semiconductor refrigeration device can be adjusted to maintain a suitable temperature for cell sheet formation within the culture vessel, thereby forming a cell sheet adsorbed onto the temperature-sensitive polymer layer. Furthermore, the temperature of the first insulating substrate of the semiconductor refrigeration device can be adjusted again to maintain a suitable temperature for separation of the cell sheet from the temperature-sensitive polymer layer, thus obtaining the cell sheet. Cell sheets obtained in this manner do not undergo enzyme or similar treatment, thus retaining the extracellular matrix secreted during cell proliferation.

[0052] Using semiconductor cooling devices to regulate the temperature of culture containers has several advantages. First, it reduces the influence of ambient temperature on the temperature of the culture container, allowing for more accurate temperature regulation. Second, semiconductor cooling devices provide rapid cooling or heating, enabling faster temperature regulation. Third, semiconductor cooling devices are easy to miniaturize, allowing cell membrane culture devices to be used in clean benches or biosafety cabinets where space is limited and cell manipulation is required.

[0053] Figure 2This is a schematic diagram illustrating the structure of a semiconductor cooling device according to one implementation of the present disclosure.

[0054] and Figure 1 Compared to the semiconductor cooling device 10 shown, Figure 2 The illustrated semiconductor cooling device 10 further includes a heat-conducting component 14. The heat-conducting component 14 is disposed on the side of the first insulating substrate 11 away from the second insulating substrate 12. For example, the heat-conducting component 14 can be in direct contact with the first insulating substrate 11. Alternatively, the heat-conducting component 14 can be bonded to the first insulating substrate 11 using thermally conductive silicone or the like to improve thermal conductivity. In some embodiments, the material of the heat-conducting component 14 may include, but is not limited to, metallic materials, polymeric materials, inorganic non-metallic materials, or composite materials of the above.

[0055] The heat-conducting component 14 facilitates the transfer of heat between the semiconductor cooling device 10 and the culture container 30, thereby enabling the semiconductor cooling device 10 to adjust the temperature of the culture container 30 more quickly.

[0056] One or more culture containers 30 may be provided on the heat-conducting component 14.

[0057] In some implementations, the culture container 30 can be directly disposed on the surface of the heat-conducting component 14.

[0058] In other implementations, such as Figure 2 As shown, the heat-conducting component 14 may have one or more grooves 141. At least one culture container 30 may be at least partially disposed in the groove 141. For example, one culture container 30 may be disposed in one groove 141. As an example, the heat-conducting component 14 may have multiple grooves 141 of different sizes, and culture containers 30 of different sizes may be disposed in the grooves 141 of different sizes. Disposing the culture container 30 in the grooves 141 of the heat-conducting component 14 can further reduce the influence of ambient temperature, thereby allowing for more precise temperature regulation of the culture container 30.

[0059] In some embodiments, the projection of the first insulating substrate 11 onto the second insulating substrate 12 lies within the projection of the heat-conducting component 14 onto the second insulating substrate 12. In this manner, the number of semiconductor thermocouples 13 and the size of the first insulating substrate 11 in the semiconductor cooling device 10 can be reduced, making the semiconductor cooling device 10 more portable.

[0060] In some embodiments, the semiconductor cooling device 10 may further include a heat dissipation component 15, such as a heat sink. The heat dissipation component 15 is disposed on the side of the second insulating substrate 12 away from the first insulating substrate 11. The heat dissipation component 15 may be in direct contact with the second insulating substrate 12, or it may be bonded to the second insulating substrate 12 by means of thermally conductive silicone or the like. The heat dissipation component 15 helps dissipate heat from the second insulating substrate 12, preventing heat accumulation on the second insulating substrate 12 from adversely affecting the performance of the semiconductor cooling device 10.

[0061] Figure 3 This is a schematic diagram illustrating the structure of a semiconductor cooling device according to another implementation of the present disclosure.

[0062] and Figure 2 Compared to the semiconductor cooling devices shown, Figure 3 The semiconductor cooling device 10 shown also includes a first switch 16, a first power supply 17, and a second power supply 18. It should be noted that, as mentioned below, one of the first and second terminals is positive and the other is negative. For example, if the first terminal is positive, the second terminal is negative, and vice versa.

[0063] The first terminal (e.g., negative terminal) of the first power supply 17 is connected to the second surface S2 of the second semiconductor portion 132 of the first semiconductor thermocouple 13 via the third metal member M3. The second terminal (e.g., positive terminal) of the first power supply 17 is configured to be connected to the second surface S2 of the first semiconductor portion 131 of the Nth semiconductor thermocouple 13 via the first switch 16 and the fourth metal member M4.

[0064] The second terminal (e.g., positive terminal) of the second power supply 18 is connected to the second surface S2 of the second semiconductor portion 132 of the first semiconductor thermocouple 13 via the third metal member M3. The first terminal (e.g., negative terminal) of the second power supply 18 is configured to be connected to the second surface S2 of the first semiconductor portion 131 of the Nth semiconductor thermocouple 13 via the first switch 16 and the fourth metal member M4.

[0065] The first switch 16 is configured to connect to either the second terminal (e.g., negative terminal) of the first power supply 17 or the first terminal (e.g., positive terminal) of the second power supply 18 in response to a user operation. In other words, depending on the user's operation, the first switch 16 can be connected to the second terminal of the first power supply 17 or to the first terminal of the second power supply 18.

[0066] When the first semiconductor section 131 is an N-type semiconductor and the second semiconductor section 132 is a P-type semiconductor: if you want to lower the temperature of the culture container 30, you can control the first switch 16 to connect to the second terminal of the first power supply 17; if you want to raise the temperature of the culture container 30, you can control the first switch 16 to connect to the first terminal of the second power supply 18.

[0067] When the first semiconductor section 131 is a P-type semiconductor and the second semiconductor section 132 is an N-type semiconductor: if you want to lower the temperature of the culture container 30, you can control the first switch 16 to connect to the first terminal of the second power supply 18; if you want to raise the temperature of the culture container 30, you can control the first switch 16 to connect to the second terminal of the first power supply 17.

[0068] In some embodiments, the first metal part M1, the second metal part M2, the third metal part M3, and the fourth metal part M4 can all be metal sheets.

[0069] In some embodiments, see Figure 3 The semiconductor cooling device 10 may also include a voltage divider adjustable element 19, such as a sliding rheostat. The voltage divider adjustable element 19 may be integrally disposed with the first switch 16. For example, the voltage divider adjustable element 19 may be a voltage divider with a switch. However, this disclosure is not limited thereto, and the voltage divider adjustable element 19 may also be disposed separately from the first switch 16.

[0070] The voltage divider adjustable element 19 is connected between the first power supply 17 and the N semiconductor thermocouples 13, and also between the second power supply 18 and the N semiconductor thermocouples 13. For example, the voltage divider adjustable element 19 can be connected between the negative terminal of the first power supply 17 and the first semiconductor thermocouple 13, and between the positive terminal of the second power supply 18 and the first semiconductor thermocouple 13. As another example, the voltage divider adjustable element 19 can be connected between the positive terminal of the first power supply 17 and the Nth semiconductor thermocouple 13, and between the negative terminal of the second power supply 18 and the Nth semiconductor thermocouple 13, such as... Figure 3 As shown.

[0071] In the above embodiments, the voltage applied between N semiconductor thermocouples can be changed by using a voltage divider adjustable element, thereby controlling the cooling or heating speed.

[0072] Figure 4 This is a schematic diagram illustrating the structure of a semiconductor cooling device according to yet another implementation of the present disclosure.

[0073] and Figure 3 Compared to the semiconductor cooling devices shown, Figure 4 The illustrated semiconductor cooling device 10 may further include a temperature monitoring component 20 and a controller 21. The temperature monitoring component 20 is configured to monitor the temperature of the first insulating substrate 11. The controller 21 is configured to control the state of the first switch 16 based on the temperature of the first insulating substrate 11. As an example, the temperature monitoring component 20 may be, for example, a bimetallic thermometer or a liquid thermometer.

[0074] For example, controller 21 can be configured to close the first switch 16 when the temperature of the first insulating substrate 11 is within a preset temperature range, i.e., connect it to one of the second poles of the first power supply 17 and the first pole of the second power supply 18. Controller 21 can also be configured to open the first switch 16 when the temperature of the first insulating substrate 11 is outside the preset temperature range, i.e., not connect it to either the second pole of the first power supply 17 or the first pole of the second power supply 18; or, control the first switch 16 to switch its connection from one of the second poles of the first power supply 17 or the first pole of the second power supply 18 to the other of the two power supplies. It should be understood that the preset temperature range can be set according to actual conditions.

[0075] Assuming the temperature of the first insulating substrate 11 is within a preset temperature range, the first switch 16 is connected to the second terminal of the first power supply 17. If the temperature of the first insulating substrate 11 is not within the preset temperature range, the controller 21 can control the first switch 16 to disconnect from the second terminal of the first power supply 17 and not connect to the first terminal of the second power supply 18, or control the first switch 16 to switch from being connected to the second terminal of the first power supply 17 to being connected to the first terminal of the second power supply 18.

[0076] Figure 5 This is a schematic diagram illustrating the structure of a semiconductor cooling device according to another implementation of the present disclosure.

[0077] and Figure 4 Compared to the semiconductor cooling device 10 shown, Figure 5 The illustrated semiconductor cooling device 10 may further include a second switch 22. In some embodiments, the second switch 22 may be integrated with the temperature monitoring component 20. The second switch 22 is connected between the first power supply 17 and N semiconductor thermocouples 13, and also connected between the second power supply 18 and N semiconductor thermocouples 13. For example, the second switch 22 may be connected between the negative terminal of the first power supply 17 and the first semiconductor thermocouple 13, and also connected between the positive terminal of the second power supply 18 and the first semiconductor thermocouple 13. As another example, the second switch 22 may be connected between the positive terminal of the first power supply 17 and the Nth semiconductor thermocouple 13, and also connected between the negative terminal of the second power supply 18 and the Nth semiconductor thermocouple 13, such as... Figure 5 As shown.

[0078] exist Figure 5 In this configuration, the controller 21 is configured to control the second switch 22 to close when the temperature of the first insulating substrate 11 is within a preset temperature range; and to control at least one of the first switch 16 and the second switch 22 to open when the temperature of the first insulating substrate 11 is outside the preset temperature range.

[0079] In some embodiments, the semiconductor cooling device 10 may further include a display configured to display the temperature of the first insulating substrate 11. Here, the temperature displayed may include Celsius, Fahrenheit, or Kelvin. In some embodiments, the display may also be configured to switch between different types of temperatures.

[0080] In some embodiments, the display can be a touch display. When using the cell membrane culture device, the user can change the state of the first switch 16 or the second switch 22, or adjust the voltage of the voltage-adjustable element, etc., through touch operation.

[0081] In other embodiments, the semiconductor cooling device 10 may also include operable components, such as knobs, buttons, etc. When using the cell membrane culture device, the user can change the state of the first switch 16 or the second switch 22, or adjust the voltage of the voltage divider adjustable element, etc., by operating the operable components. For example, one knob can be operated to connect the first switch 16 to the second terminal of the first power supply, and another knob can be operated to connect the first switch 16 to the first terminal of the second power supply.

[0082] Figure 6 This is a schematic flowchart illustrating a method for preparing a cell membrane sheet according to an embodiment of the present disclosure. The method for preparing the cell membrane sheet can be implemented based on the cell membrane sheet culture apparatus of any of the above embodiments.

[0083] like Figure 6 As shown, the method for preparing cell membrane sheets may include steps 602 and 604.

[0084] In step 602, the temperature of the first insulating substrate of the semiconductor cooling device is adjusted to a first temperature so that multiple cells in the culture container form cell membranes adsorbed onto the temperature-sensitive polymer layer.

[0085] Here, multiple cells can be from mammals, birds, etc. The initial temperature can be adjusted according to the cell type. For example, the initial temperature range can be approximately 37°C to approximately 38°C.

[0086] In some embodiments, a temperature-sensitive polymer layer is disposed at the bottom of the culture container. In this case, a culture medium containing multiple cells can be directly applied to the culture container, and then the temperature of the first insulating substrate of the semiconductor cooling device can be adjusted to a first temperature.

[0087] In other embodiments, the bottom of the culture container is not provided with a temperature-sensitive polymer layer. In this case, a temperature-sensitive polymer layer can be formed at the bottom of the culture container first, then a culture medium containing multiple cells can be applied to the culture container, and then the temperature of the first insulating substrate of the semiconductor cooling device can be adjusted to a first temperature.

[0088] In step 604, the temperature of the first insulating substrate is adjusted to a second temperature different from the first temperature, so that the cell membrane sheet separates from the temperature-sensitive polymer layer.

[0089] In some embodiments, the second temperature may be lower than the first temperature. For example, the range of the second temperature may be about 15°C to about 25°C.

[0090] In the above embodiments, semiconductor cooling devices are used to regulate the temperature of the culture container. On the one hand, this reduces the influence of ambient temperature on the temperature of the culture container, making the temperature regulation of the culture container more accurate. On the other hand, semiconductor cooling devices have a fast cooling or heating speed, making the temperature regulation of the culture container faster.

[0091] The embodiments of this disclosure have now been described in detail. To avoid obscuring the concept of this disclosure, some details known in the art have not been described. Those skilled in the art can fully understand how to implement the technical solutions disclosed herein based on the above description.

[0092] While specific embodiments of this disclosure have been described in detail by way of examples, those skilled in the art should understand that the examples are for illustrative purposes only and not intended to limit the scope of this disclosure. Those skilled in the art should understand that modifications can be made to the above embodiments or equivalent substitutions can be made to some technical features without departing from the scope and spirit of this disclosure. The scope of this disclosure is defined by the appended claims.

Claims

1. A cell membrane sheet culture device, comprising: Semiconductor cooling devices, including: First insulating substrate. Second insulating substrate, A heat-conducting component is disposed on the side of the first insulating substrate away from the second insulating substrate. The heat-conducting component has multiple grooves, and At least one semiconductor thermocouple is disposed between the first insulating substrate and the second insulating substrate, wherein the at least one semiconductor thermocouple comprises N semiconductor thermocouples, where N is an integer greater than or equal to 2, wherein: Each semiconductor thermocouple includes a first semiconductor portion and a second semiconductor portion with different conductivity types. Each of the first and second semiconductor portions includes a first surface and a second surface disposed opposite to each other. The first surface is closer to the first insulating substrate than the second surface. The first surfaces of the first semiconductor portion and the second semiconductor portion are connected by a first metal member. The second surface of the first semiconductor portion of the i-th semiconductor thermocouple and the second surface of the second semiconductor portion of the (i+1)-th semiconductor thermocouple are connected by a second metal component, where 1 ≤ i ≤ N-1; and Multiple culture containers are configured as cell culture sheets and are separately disposed from the semiconductor cooler, with one culture container disposed in a recess; The semiconductor cooling device further includes a first switch, a first power supply, a second power supply, and a voltage divider adjustable element, wherein: The first pole of the first power supply is connected to the second surface of the second semiconductor portion of the first semiconductor thermocouple via a third metal component, and the second pole of the first power supply is configured to be connected to the second surface of the first semiconductor portion of the Nth semiconductor thermocouple via the first switch and a fourth metal component. The second pole of the second power supply is connected to the second surface of the second semiconductor portion of the first semiconductor thermocouple via the third metal member, and the first pole of the second power supply is configured to be connected to the second surface of the first semiconductor portion of the Nth semiconductor thermocouple via the first switch and the fourth metal member. The first switch is configured to connect to one of the second pole of the first power supply and the first pole of the second power supply in response to a user operation; The voltage divider adjustable element is connected between the first power supply and N semiconductor thermocouples, and is also connected between the second power supply and N semiconductor thermocouples. The voltage divider adjustable element and the first switch are integrally configured. Wherein, one of the first electrode and the second electrode is a positive electrode and the other is a negative electrode; the semiconductor cooling device further includes: A temperature monitoring component is configured to monitor the temperature of the first insulating substrate; The second switch is connected between the first power source and the N semiconductor thermocouples, and is also connected between the second power source and the N semiconductor thermocouples. The controller is configured to close the first switch and the second switch when the temperature of the first insulating substrate is within a preset temperature range, and to open the first switch and the second switch when the temperature of the first insulating substrate is outside the preset temperature range.

2. The cell sheet culture device according to claim 1, wherein The projection of the first insulating substrate onto the second insulating substrate is within the projection of the heat-conducting component onto the second insulating substrate.

3. The cell sheet culture device according to claim 1, wherein At least one of the culture containers has a temperature-sensitive polymer layer disposed on the bottom side away from the first insulating substrate, and the surface energy of the temperature-sensitive polymer layer changes monotonically with temperature changes.

4. The cell sheet culture device according to claim 3, wherein The surface energy of the temperature-sensitive polymer layer increases with increasing temperature.

5. The cell sheet culture device according to claim 1, wherein The semiconductor cooling device further includes: A heat dissipation component is disposed on the side of the second insulating substrate away from the first insulating substrate.

6. A method for preparing a cell membrane sheet, said preparation method being based on the cell membrane sheet culture apparatus according to any one of claims 1-5; The preparation method includes: The temperature of the first insulating substrate of the semiconductor cooling device is adjusted to a first temperature so that multiple cells in the culture container form cell membranes adsorbed on the temperature-sensitive polymer layer. and The temperature of the first insulating substrate is adjusted to a second temperature different from the first temperature, so that the cell membrane sheet separates from the temperature-sensitive polymer layer.

7. The method of claim 6, wherein the cell sheet is prepared by the method of claim 1. The second temperature is lower than the first temperature.

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