A method for manufacturing a near-infrared light responsive MXene hydrogel film intelligent cell culture container

Abstract

The application provides a preparation method of a near-infrared light responsive MXene hydrogel film intelligent cell culture container, and solves the problems of complex preparation process, long cycle, high cost, low efficiency, poor biocompatibility and poor stability of the existing near-infrared light responsive cell culture container. The hydrophilic treatment of the cell culture container (substrate) increases the number of hydroxyl groups on the surface, thereby increasing the bonding force between the MXene hydrogel film and the cell culture container, making it more stable. First, a relatively mild method is used to synthesize MXene suspension. MXene has good photothermal effect. Then, the MXene is stirred uniformly with low-melting-point agarose and polyvinyl alcohol (PVA) which has good biocompatibility. A certain thickness of film is obtained on the cell culture container by spin coating, and the final product is obtained by using the cycle freezing and thawing.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of responsive cell culture containers, and particularly relates to a method for manufacturing a near-infrared light responsive MXene hydrogel film intelligent cell culture container. BACKGROUND

[0002] For adherent cells, detaching and transferring them from the wall surface of a cell culture plate, a culture bottle or a culture dish is one of the most basic and most commonly involved experimental operations in cell engineering and tissue engineering. The commonly used cell detaching method is trypsin digestion. Trypsin is a kind of protease, which plays a role of digestive enzyme in the body of vertebrates. Trypsin hydrolyzes the extracellular matrix protein (ECM) between cells and between cells and the surface of a culture container, thereby separating cells, and thus can be used to separate cells in tissues and detach adherent cells in the process of animal cell culture. However, in addition to the ability to digest the intercellular matrix protein, long-term action of trypsin can also digest cell membrane proteins, which has a damaging effect on cells. Therefore, when using trypsin to digest cells, it is necessary to strictly control the concentration, action time, temperature and flushing frequency of trypsin and other parameters. Under the conditions of weak alkalinity and 37℃ temperature, the digestion ability of trypsin solution is the strongest, but a slight negligence in operation can easily cause damage to the membrane structure and functional integrity of cells, thereby causing a decrease in cell activity, and even leading to cell death.

[0003] In view of the above problems, in the past few decades, a variety of methods have been developed to regulate cell behavior, mainly including mechanical scratching, temperature control, pH control, ion control, magnetization control and light control. Light control method is outstanding among many separation technologies due to its strong universality, fast separation speed and good controllability. In the light control method, near-infrared light is more preferred because it causes the least damage to cells. The core of the near-infrared light control method is the design and construction of a photothermal surface, which is a surface embedded or coated with a photothermal agent such as noble metal nanomaterials, semiconductor nanomaterials, carbon-based nanomaterials and conjugated polymers. The surface can absorb light of appropriate wavelength (usually in the near-infrared region) and convert light energy to heat energy, so as to detach the cell sheet.

[0004] MXene can be used as a plasmonic substrate to convert light energy into heat energy through plasmonic resonance. Plasmonic resonance refers to the excited oscillation of electrons under external illumination when the light frequency coincides with the oscillation frequency of the electrons. MXene is a common plasmonic material because it has a large electron density. Currently, the main method for modifying MXene on the surface of a cell culture plate is solution casting polymerization. Solution casting polymerization mainly uses surface binding force to adhere MXene to the surface of a cell culture plate (see Zhao Y, Yan C, Hou T, et al. Multifunctional Ti3C2T x MXene-BasedComposite Coatings with Superhydrophobic Anti-icing and Photothermal DeicingProperties[J].ACS Applied Materials&Interfaces,2022,14(22):26077-26087.)。

[0005] Although the above-mentioned methods can achieve controllable detachment of cells, there are still the following disadvantages in using the above-mentioned methods: the solution casting polymerization method also has strict requirements for the surface properties of the substrate. Generally, the surface of the substrate needs to be treated complicatedly to generate some groups on the surface of the substrate to increase the surface binding force, and then a thin film is directly polymerized on the surface. The thin film and the substrate are connected by hydrogen bond van der Waals force, etc., so that the thin film and the substrate surface are tightly adhered together. However, the binding force is weak, and the use stability of the modified cell culture plate is poor. These problems not only increase the production cost of the near-infrared light responsive cell culture plate, but also greatly reduce the production efficiency.

[0006] Therefore, it is of great significance to develop and design a simple, inexpensive and efficient method to produce a near-infrared light responsive intelligent cell culture container with good use stability, which promotes the market development of the near-infrared light responsive cell culture container. SUMMARY

[0007] The purpose of the present application is to solve the problems of complex production process, long cycle, high cost, low efficiency, poor biocompatibility and poor stability of the existing near-infrared light responsive cell culture container, and to provide a production method of a near-infrared light responsive MXene hydrogel film intelligent cell culture container.

[0008] To achieve the above-mentioned purpose, the technical solution provided by the present application is:

[0009] A preparation method of a near-infrared light responsive MXene hydrogel film, characterized by comprising the following steps:

[0010] 1) mixing concentrated hydrochloric acid and lithium fluoride, stirring to obtain a hydrofluoric acid solution; the hydrofluoric acid synthesized by this method has less toxicity and the preparation environment is friendly;

[0011] 2) mixing the hydrofluoric acid solution obtained in step 1) with Ti3AlC2, and then stirring and heating to obtain a mixture;

[0012] 3) washing the mixture obtained in step 2) for multiple times (3-5 times), and centrifuging to remove the supernatant to obtain Ti3C2T x precipitate;

[0013] 4) dispersing the Ti3C2T x precipitate obtained in step 3) into water, and then ultrasonicating and centrifuging under an inert gas (such as argon) to collect the supernatant to obtain a MXene suspension, i.e., a MXene nanosheet-containing suspension;

[0014] 5) mixing the MXene suspension obtained in step 4) with low-melting agarose and PVA powder, and then stirring and heating to obtain a mixed solution;

[0015] 6) pouring the mixed solution obtained in step 5) onto a substrate that has been subjected to surface hydrophilic treatment, and then sequentially performing spin coating, freezing, thawing, and cyclic operation multiple times to prepare a near-infrared light-responsive MXene hydrogel film on the substrate; in this step, the cyclic freezing and thawing is used to obtain a more uniform and better-performing film. During freezing, the PVA chains form hydrogen bonds through hydroxyl groups, and the polymer chains are locally ordered to form high-molecular crystals. Through cyclic freezing and thawing, the crystallinity of PVA is continuously enhanced, thereby forming an opaque hydrogel with crystal regions as physical crosslinking points.

[0016] Further, in step 1), the molar concentration of the concentrated hydrochloric acid is 9M, i.e., the molar concentration of the commercially available product.

[0017] The mass ratio of the concentrated hydrochloric acid to lithium fluoride is 15-30:1; the stirring time is 1-60 min, preferably 1-10 min.

[0018] Further, in step 2), the feeding ratio of Ti3AlC2 to the hydrofluoric acid solution is 1:1-50, g / mL, preferably 1:15-25, g / mL; the stirring temperature is 20-100℃, preferably 35-50℃; and the time is 10-100 h, preferably 12-24 h.

[0019] Further, in step 3), the deionized water is used for washing until the pH of the supernatant is 5-7, preferably 6; the centrifugation speed is 500-10000 rpm, preferably 3000-5000 rpm; and the time is 1-60 min, preferably 5-10 min.

[0020] Further, in step 4), the mass ratio of Ti3C2T x The ratio of the precipitate and water is 1:1-50 g / mL, preferably 1:15-25 g / mL; the ultrasonic time is 10-100 min, preferably 20-40 min; the centrifugal speed is 500-10000 rpm, preferably 3000-5000 rpm, and the time is 1-200 min, preferably 60-80 min.

[0021] Further, in step 5), the mass ratio of the MXene suspension, low-melting agarose and PVA powder is 10-100:1-50:1-50, preferably 10-20:1:3-5;

[0022] The heating temperature is 40-150℃, preferably 80-90℃, and the stirring time is 1-12h, preferably 2-4h;

[0023] In step 6), the spin coating machine is used to spin coat at a speed of 500-3000 r / min (preferably 1000-3000 r / min) for 1-5 min, and then freeze it at-80℃ to-20℃ for 1-24h (preferably 3-8h), thaw it at 10-40℃ (preferably 10-30℃) for 0.5-12h (preferably 1-3h), and repeat the cycle 2-10 times (preferably 2-5 times).

[0024] The application provides a near-infrared light responsive MXene hydrogel film, which is prepared by the above method and has a thickness of 50-100 μm.

[0025] The near-infrared light responsive MXene hydrogel film prepared by the above method is used as a photothermal surface in the preparation of a near-infrared light responsive intelligent cell culture container.

[0026] In addition, the application also provides a preparation method of a near-infrared light responsive intelligent cell culture container, which is characterized by: preparing a near-infrared light responsive MXene hydrogel film on a cell culture container with a hydrophilic surface treatment according to the above method to obtain a near-infrared light responsive intelligent cell culture container; that is, the cell culture container is directly used as a substrate, a near-infrared light responsive MXene hydrogel film is modified thereon and used as a photothermal surface, which is beneficial to cell detachment and transfer.

[0027] Further, the cell culture container is a cell culture plate, a cell culture bottle or a cell culture dish, and the material of the cell culture container can be any material, for example, the commercialized cell culture containers with existing glass, polystyrene, polypropylene, polyvinyl chloride and polyethylene materials are all applicable.

[0028] And a near-infrared light response type intelligent cell culture container, which is characterized in that: the preparation method is used.

[0029] The application of the above-mentioned near-infrared light response type intelligent cell culture container in cell sheet detachment and separation.

[0030] The principle of the application:

[0031] The hydrophilic treatment of the cell culture container (substrate) in the application increases the number of hydroxyl groups on the surface, thereby increasing the bonding force between the MXene hydrogel film and the cell culture container, making it more stable. First, a relatively mild method is used to synthesize MXene suspension. MXene has good photothermal effect. Then, it is stirred together with low-melting-point agarose and polyvinyl alcohol (PVA) with good biocompatibility. A 50-100 mu m thick film is obtained on the cell culture container by spin coating, and the final product is obtained by cyclic freezing and thawing. Cells are cultured on the MXene hydrogel film. The MXene layer has near-infrared response function, which can heat the cell layer by absorbing near-infrared light. When the temperature reaches a certain value, the low-melting-point agarose in the MXene hydrogel film dissolves, and the cell layer separates from the MXene hydrogel film layer, thereby realizing complete separation.

[0032] Advantages of the application:

[0033] 1. The MXene nanosheet prepared by etching and stripping method is mixed with low-melting-point agarose and PVA powder to prepare MXene hydrogel. Under near-infrared light irradiation, the MXene hydrogel can reach equilibrium temperature within 200 seconds. Due to its excellent anti-swelling ability and near 100% photothermal conversion efficiency, it shows great potential in near-infrared light response. It can be seen that the MXene film prepared by the application has higher photothermal conversion efficiency and can reach thermal equilibrium faster. The main reason is that the Mxene suspension obtained by the method of the application is more uniform than the suspension obtained by processing the purchased Mxene solid, so the distribution of MXene in the film is also more uniform and stable.

[0034] 2. The MXene hydrogel prepared by the application has better biocompatibility. PVA and low-melting-point agarose have excellent biocompatibility. By mixing PVA, low-melting-point agarose and MXene together, the biocompatibility can be greatly improved.

[0035] 3. The application uses oxygen plasma treatment method to hydrophilic treatment of the cell culture container (substrate), so that the MXene hydrogel film and the substrate surface are combined by hydrogen bond (chemical bond), which has good stability and is more stable.

[0036] 4. The method of the present application for modifying the near-infrared light responsive MXene hydrogel film on the cell culture container has simple process steps, short cycle, high efficiency, widely available raw materials, and lower cost than existing methods. BRIEF DESCRIPTION OF DRAWINGS

[0037] Figure 1 is an optical image of the MXene hydrogel film cell culture dish prepared in Example 1;

[0038] Figure 2 is an optical microscope image of the culture of human renal clear cell adenocarcinoma cells using the MXene hydrogel film cell culture dish in Example 1;

[0039] Figure 3 is the photothermal response desorption efficiency of human renal clear cell adenocarcinoma cells by the MXene hydrogel film cell culture dish in Example 1;

[0040] Figure 4 is the photothermal response desorption efficiency of human renal clear cell adenocarcinoma cells by the MXene hydrogel film cell culture dish in Example 1 in the second reuse;

[0041] Figure 5 is the cell activity test result of the culture of cells using the MXene hydrogel film cell culture dish in Example 1 for 24h and 72h. DETAILED DESCRIPTION

[0042] The content of the present application is further described in detail below in combination with the drawings and specific examples:

[0043] Unless otherwise specified, the present application has no special requirements for the source of the raw materials used, and commercially available products known to those skilled in the art can be used.

[0044] The present application provides a method for preparing a near-infrared light responsive intelligent cell culture container, comprising the following steps:

[0045] Step 1): Mix concentrated hydrochloric acid and lithium fluoride in a Teflon beaker and stir for 1-60 min to obtain a hydrofluoric acid solution; the molar concentration of concentrated hydrochloric acid is 9M, and the mass ratio of concentrated hydrochloric acid to lithium fluoride is 15-30:1;

[0046] Step 2): Mix the hydrofluoric acid solution obtained in step 1) with Ti3AlC2 and heat and stir for 10-100h to obtain a mixture; the stirring temperature is 20-100℃; the feeding ratio of Ti3AlC2 to hydrofluoric acid solution is 1:1-50, g / mL;

[0047] Step 3): The mixture obtained in step 2) is washed with deionized water for 3-5 times until the supernatant pH = 6, and then centrifuged for 1-60 min to remove the supernatant to obtain Ti3C2T x precipitate; the centrifugal speed is 500-10000 rpm;

[0048] Step 4): The Ti3C2T x precipitate obtained in step 3) is redispersed in deionized water, ultrasonicated for 10-100 min under an argon atmosphere, and then centrifuged for 1-200 min to collect the supernatant to obtain a MXene suspension; the centrifugal speed is 500-10000 rpm; the mass ratio of Ti3C2T x precipitate to water is 1:1-50, g / mL;

[0049] Step 5): The MXene suspension obtained in step 4) is mixed with low-melting agarose and PVA powder, and heated and stirred; the heating temperature is 40-150°C, and the stirring time is 1-12 h; wherein the mass ratio of MXene suspension, low-melting agarose and PVA powder is 10-100:1-50:1-50;

[0050] Step 6): The mixed solution obtained in step 5) is poured into a cell culture container whose surface has been hydrophilically treated, and is spin-coated at a speed of 500-3000 r / min for 1-5 min using a spin coater, frozen for 1-24 h, thawed for 0.5-12 h, and cycled 2-10 times to prepare a near-infrared light-responsive intelligent cell culture container; wherein the freezing temperature is -80°C to -20°C, and the thawing temperature is 10-40°C.

[0051] The above process is used to manufacture a near-infrared light-responsive intelligent cell culture container, and the process parameters of a specific example are as follows:

[0052] Example 1

[0053] Step 1): 20 mL of 9M hydrochloric acid and 1 g of lithium fluoride are mixed in a Teflon beaker and stirred for 5 min to obtain a hydrofluoric acid solution;

[0054] Step 2): The hydrofluoric acid solution obtained in step 1) is mixed with 1 g of Ti3AlC2, and stirred at 35°C for 24 h to obtain a mixture;

[0055] Step 3): The mixture obtained in step 2) is washed with deionized water until the supernatant pH = 6, and then centrifuged at 3500 rpm for 5 min to remove the supernatant to obtain Ti3C2T x precipitate;

[0056] Step 4): The Ti3C2Tx The precipitate was re-dispersed into 20 mL of deionized water, ultrasonic treatment was performed for 20 min under argon atmosphere, and then centrifugation was performed at 3500 rpm for 1 h, the supernatant was collected to obtain a MXene suspension;

[0057] Step 5): The MXene suspension obtained in step 4) was mixed with 0.3 g of low-melting-point agarose and 1 g of PVA powder, and heated and stirred; stirring was performed at 90°C for 2 h to obtain a uniform mixture.

[0058] Step 6): The uniform mixture obtained in step 5) was poured into a cell culture container whose surface had been subjected to hydrophilic treatment, and spin coating was performed at a speed of 1000 r / min for 3 min using a spin coater, freezing was performed at -20°C for 8 h, thawing was performed at 25°C for 2 h, and the cycle was repeated for 3 times, to finally obtain a near-infrared light-responsive intelligent cell culture container.

[0059] Example 2

[0060] Step 1): 15 mL of 9M hydrochloric acid and 1 g of lithium fluoride were mixed in a Teflon beaker, and stirred for 1 min to obtain a hydrofluoric acid solution;

[0061] Step 2): The hydrofluoric acid solution obtained in step 1) was mixed with 0.5 g of Ti3AlC2, and stirring was performed at 35°C for 12 h to obtain a mixture;

[0062] Step 3): The mixture obtained in step 2) was washed with deionized water until the supernatant pH = 6, and then centrifugation was performed at 5000 rpm for 10 min, the supernatant was removed, and Ti3C2T x precipitate was obtained;

[0063] Step 4): The Ti3C2T x precipitate obtained in step 3) was re-dispersed into 20 mL of deionized water, ultrasonic treatment was performed for 20 min under argon atmosphere, and then centrifugation was performed at 3000 rpm for 1 h, the supernatant was collected to obtain a MXene suspension;

[0064] Step 5): The MXene suspension obtained in step 4) was mixed with 0.1 g of low-melting-point agarose and 0.3 g of PVA powder, and heated and stirred; stirring was performed at 90°C for 2 h to obtain a uniform mixture.

[0065] Step 6): The uniform mixture obtained in step 5) was poured into a cell culture container whose surface had been subjected to hydrophilic treatment, and spin coating was performed at a speed of 3000 r / min for 5 min using a spin coater, freezing was performed at -20°C for 3 h, thawing was performed at 10°C for 1 h, and the cycle was repeated for 2 times, to finally obtain a near-infrared light-responsive intelligent cell culture container.

[0066] Example 3

[0067] Step 1): 30 mL of 9M hydrochloric acid and 1 g of lithium fluoride were mixed in a Teflon beaker, stirred for 10 min to obtain a hydrofluoric acid solution;

[0068] Step 2): The hydrofluoric acid solution obtained in step 1) was mixed with 2 g of Ti3AlC2, and stirred at 50°C for 24 h to obtain a mixture;

[0069] Step 3): The mixture obtained in step 2) was washed with deionized water until the supernatant pH = 6, then centrifuged at 3000 rpm for 5 min, the supernatant was removed, and Ti3C2T x precipitate was obtained;

[0070] Step 4): The Ti3C2T x precipitate obtained in step 3) was re-dispersed in 20 mL of deionized water, ultrasonicated for 40 min in an argon environment, then centrifuged at 5000 rpm for 1.3 h, and the supernatant was collected to obtain a MXene suspension;

[0071] Step 5): The MXene suspension obtained in step 4) was mixed with 0.5 g of low-melting agarose and 1.5 g of PVA powder, and heated and stirred; the mixture was stirred at 90°C for 2 h to obtain a uniform mixture.

[0072] Step 6): The uniform mixture obtained in step 5) was poured into a cell culture container whose surface had been hydrophilically treated, and was spin-coated at a speed of 1000 r / min for 1 min using a spin coater, then frozen at -80°C for 8 h, thawed at 30°C for 2 h, and the cycle was repeated 5 times to finally obtain a near-infrared light responsive intelligent cell culture container.

[0073] Example 4

[0074] Step 1): 12 mL of 9M hydrochloric acid and 1 g of lithium fluoride were mixed in a Teflon beaker, stirred for 2 min to obtain a hydrofluoric acid solution;

[0075] Step 2): The hydrofluoric acid solution obtained in step 1) was mixed with 0.8 g of Ti3AlC2, and stirred at 35°C for 18 h to obtain a mixture;

[0076] Step 3): The mixture obtained in step 2) was washed with deionized water until the supernatant pH = 6, then centrifuged at 4000 rpm for 5 min, and the supernatant was removed to obtain Ti3C2T x precipitate;

[0077] Step 4): The Ti3C2T xThe precipitate was re-dispersed into 20 mL of deionized water, ultrasonic treatment was performed for 20 min under an argon atmosphere, and then centrifugation was performed at 3000 rpm for 1 h, the supernatant was collected, and a MXene suspension was obtained;

[0078] Step 5): The MXene suspension obtained in step 4) was mixed with 0.2 g of low-melting-point agarose and 1 g of PVA powder, and heated and stirred; stirring was performed at 90°C for 2 h to obtain a uniform mixture.

[0079] Step 6): The uniform mixture obtained in step 5) was poured into a cell culture container whose surface had been subjected to hydrophilic treatment, and spin coating was performed at a speed of 3000 r / min for 5 min using a spin coater, freezing was performed at -50°C for 5 h, thawing was performed at 30°C for 1 h, and the cycle was repeated 5 times, and finally a near-infrared light-responsive intelligent cell culture container was obtained.

[0080] Example 5

[0081] Step 1): 25 mL of 9M hydrochloric acid and 1 g of lithium fluoride were mixed in a Teflon beaker, and stirring was performed for 8 min to obtain a hydrofluoric acid solution;

[0082] Step 2): The hydrofluoric acid solution obtained in step 1) was mixed with 1.5 g of Ti3AlC2, and stirring was performed at 45°C for 24 h to obtain a mixture;

[0083] Step 3): The mixture obtained in step 2) was washed with deionized water until the supernatant pH was 6, and then centrifugation was performed at 3000 rpm for 5 min, the supernatant was removed, and Ti3C2T x precipitate;

[0084] Step 4): The Ti3C2T x precipitate obtained in step 3) was re-dispersed into 20 mL of deionized water, ultrasonic treatment was performed for 30 min under an argon atmosphere, and then centrifugation was performed at 4000 rpm for 1.2 h, the supernatant was collected, and a MXene suspension was obtained;

[0085] Step 5): The MXene suspension obtained in step 4) was mixed with 0.4 g of low-melting-point agarose and 1 g of PVA powder, and heated and stirred; stirring was performed at 90°C for 2 h to obtain a uniform mixture.

[0086] Step 6): The uniform mixture obtained in step 5) was poured into a cell culture container whose surface had been subjected to hydrophilic treatment, and spin coating was performed at a speed of 1000 r / min for 1 min using a spin coater, freezing was performed at -40°C for 4 h, thawing was performed at 30°C for 1 h, and the cycle was repeated 5 times, and finally a near-infrared light-responsive intelligent cell culture container was obtained.

[0087] Figure 1The optical image of the MXene hydrogel film cell culture dish (i.e. the near-infrared light responsive intelligent cell culture dish) made for Example 1 can clearly show that the cell culture dish has an opaque film on the bottom. After that, human renal clear cell adenocarcinoma cells were cultured on the cell culture dish as shown in Figure 1 The cell sheet can be obtained as shown in Figure 2 It can be determined that the MXene hydrogel film cell culture dish made by the present application can successfully carry out cell culture.

[0088] In order to verify the near-infrared light response ability and stability (reusability) of the MXene hydrogel film cell culture dish made by the present application, the present application also uses near-infrared light to irradiate the culture dish to carry out the desorption test, and the desorption efficiency is as shown in Figure 3 It can be seen that the MXene hydrogel film cell culture dish made by the present application has good near-infrared light response ability, and converts light energy into heat energy to make the cell sheet desorb; after washing the culture dish on which the desorbed cell sheet is placed, the culture dish is used to continue to culture the cell sheet and carry out the desorption process, and the desorption efficiency of the second desorption of the cell sheet is as shown in Figure 4 It can be seen that the desorption ability is still high when the culture dish is used again, and the stability is good. In addition, the biocompatibility of the MXene hydrogel film cell culture dish made by the present application is detected, and the cell activity detection is carried out for 24h and 72h, and the detection results are as shown in Figure 5 It can be seen that the cell culture dish made by the present application has good biocompatibility.

[0089] In summary, the cell culture dish made by the present application can carry out intelligent response under near-infrared light irradiation, has simple manufacturing process, low cost, high efficiency, good biocompatibility and stability.

[0090] The above is only a specific embodiment of the present application, but the protection scope of the present application is not limited thereto, and any skilled person in the art can easily think of various equivalent modifications or replacements within the technical range disclosed by the present application, and these modifications or replacements should be covered within the protection scope of the present application.

Claims

1. A method for manufacturing a near-infrared light-responsive intelligent cell culture container, characterized by: A near-infrared light-responsive MXene hydrogel film is prepared on a cell culture container with a hydrophilic surface treatment to obtain a near-infrared light-responsive smart cell culture container. The near-infrared light-responsive MXene hydrogel film was prepared according to the following method: 1) Mix concentrated hydrochloric acid and lithium fluoride, stir and react to obtain a hydrofluoric acid solution; 2) mixing the hydrofluoric acid solution obtained in step 1) with Ti3AlC2, heating and stirring to react, to obtain a mixture; 3) The mixture obtained in step 2) was washed several times and centrifuged to remove the supernatant to obtain Ti3C2T x sediment; 4) Ti3C2T obtained in step 3) x The precipitate was redispersed in water, sonicated and then centrifuged under an inert gas environment, and the supernatant was collected to obtain a MXene suspension; 5) mixing the MXene suspension obtained in step 4) with low melting point agarose and polyvinyl alcohol powder, heating and stirring to obtain a mixed solution; 6) The mixed solution obtained in step 5) is inverted onto a cell culture container with a hydrophilic surface treatment, and spin coating, freezing, and thawing operations are performed in sequence, and the cycle is repeated multiple times to prepare a MXene hydrogel film on the cell culture container.

2. The production method according to claim 1, characterized in that: In step 1), the molar concentration of the concentrated hydrochloric acid is 9 M; The mass ratio of concentrated hydrochloric acid to lithium fluoride is 15-30:1, and the reaction is stirred for 1-60 minutes.

3. The production method according to claim 1 or 2, characterized in that: In step 2), the feed ratio of Ti3AlC2 to hydrofluoric acid solution is 1:1~50g / mL; The stirring temperature is 20~100℃ and the time is 10~100h.

4. The production method according to claim 3, characterized in that: In step 3), washing with deionized water until the pH of the supernatant is 5-7; The centrifugal speed is 500-10000 rpm, and the time is 1-60 min; In step 4), Ti3C2T x The ratio of sediment to water is 1:1~50g / mL; The ultrasonic time is 10 to 100 minutes; The centrifugal speed is 500-10000 rpm, and the time is 1-200 min.

5. The production method according to claim 4, characterized in that: In step 5), the mass ratio of MXene suspension, low melting point agarose and polyvinyl alcohol powder is 10-100:1-50:1-50; The heating temperature is 40~150℃ and the stirring time is 1~12h; In step 6), the spin coater is used to spin coat the substrate at a speed of 500-3000 r / min for 1-5 minutes, and then the substrate is frozen at -80°C to -20°C for 1-24 hours and thawed at 10-40°C for 0.5-12 hours, and this cycle is repeated 2-10 times.

6. A near-infrared light-responsive intelligent cell culture container, characterized by: The method is obtained by any one of claims 1 to 5.

7. Use of the near-infrared light-responsive intelligent cell culture container according to claim 6 in the desorption and separation of cell sheets.

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