Preparation process of cell culture fluid

Abstract

The invention relates to the field of cell culture, in particular to a process for preparing a cell culture solution. Comprising the following steps: S1, selecting a basic culture medium, and adding an additive into the culture medium; s2, a culture medium and an additive are added into the preparation device, so that the culture medium and the additive are dissolved in the deionized water; s3, enabling the dissolved culture medium to flow from the upper part of the preparation device to the lower part of the preparation device, filtering the prepared culture solution through a filter membrane, removing possible microorganisms and impurities, and ensuring the sterility of the culture solution; s4, subpackaging the filtered culture solution into sterile culture bottles, and sterilizing the culture bottles by using a high-pressure steam sterilizer or a dry heat sterilizer to kill residual microorganisms; and S5, adjusting the pH value of the culture solution by using an acid or alkali solution. The prepared culture solution can be filtered through the filter membrane, possible microorganisms and impurities are removed, and the sterility of the culture solution is ensured.

Description

Technical Field

[0001] This invention relates to the field of cell culture, and more specifically to a process for preparing cell culture medium. Background Technology

[0002] Cell culture medium is a liquid culture medium used for cell culture, containing the nutrients and growth factors necessary for cell growth. Cell culture medium typically consists of various components, including inorganic salts, amino acids, sugars, vitamins, and fetal bovine serum. The formulation of cell culture medium varies depending on the type of cell and the culture purpose; a suitable medium can be selected for specific cell types to promote cell growth and proliferation. Cell culture medium needs to be prepared and stored under sterile conditions to ensure cell health and successful culture. Traditional cell culture medium preparation processes can lead to bacterial contamination, making it impossible to filter the prepared medium through a membrane, thus failing to remove potential microorganisms and impurities and compromising the sterility of the medium. Summary of the Invention

[0003] To overcome the shortcomings of the prior art, the present invention provides a cell culture medium preparation process, which has the advantage of filtering the prepared culture medium through a filter membrane to remove any possible microorganisms and impurities, thus ensuring the sterility of the culture medium.

[0004] A cell culture medium preparation process includes the following steps:

[0005] S1: Select a basic culture medium and add additives to the culture medium;

[0006] S2: Add the culture medium and additives to the preparation device so that the culture medium and additives dissolve in deionized water;

[0007] S3: Allow the dissolved culture medium to flow from the top to the bottom of the preparation device, filter the prepared culture solution through a filter membrane to remove any possible microorganisms and impurities, and ensure the sterility of the culture solution;

[0008] S4: Dispense the filtered culture medium into sterile culture bottles and sterilize the culture bottles using an autoclave or dry heat sterilizer to kill any remaining microorganisms.

[0009] S5: Adjust the pH of the culture medium using acid or alkaline solutions. Attached Figure Description

[0010] The present invention will now be described in further detail with reference to the accompanying drawings and specific implementation methods.

[0011] Figure 1 A flowchart of a cell culture medium preparation process;

[0012] Figure 2 Flow chart for Example 1

[0013] Figure 3 Flow chart for Example 2

[0014] Figure 4 Structure diagram of the preparation device Figure 1 ;

[0015] Figure 5 Structure diagram of the preparation device Figure 2 ;

[0016] Figure 6 Structure diagram of the preparation device Figure 3 ;

[0017] Figure 7 Structure diagram of the preparation device Figure 4 ;

[0018] Figure 8 Structure diagram of the round box and the L-shaped frame

[0019] Figure 9 Structure diagram of the round box

[0020] Figure 10 Structure diagram of the bottom box

[0021] Figure 11 Structure diagram of the bottom box and the vertical frame

[0022] Figure 12 Structure diagram of the conical plate Figure 1 ;

[0023] Figure 13 Structure diagram of the conical plate Figure 2 .

[0024] In the figure: round box 101; middle cylinder 102; back seat 103;

[0025] L-shaped frame 201; vertical shaft 202; motor 203; gear 204; rotating frame 205; smearing frame 206; tab 207; toothed cylinder 208; pressing plug 209;

[0026] Bottom box 301; filter membrane 302; middle rod 303; filter plate 304; insertion rod 305; insertion seat 306;

[0027] Vertical frame 401; sliding sleeve 402; square column 403; round sheet 404; clamping groove 405;

[0028] Conical plate 501; blocking sheet 502; L-shaped hole 503; round seat 504. DETAILED DESCRIPTION

[0029] Example 1:

[0030] A cell culture solution preparation process, comprising the following steps:

[0031] S1: selecting a basic culture medium, and adding additives to the culture medium;

[0032] S2: adding the culture medium and the additives into a preparation device, so that the culture medium and the additives are dissolved in deionized water;

[0033] S3: making the dissolved culture medium flow from the upper part to the lower part of the preparation device, filtering the prepared culture solution through a filter membrane to remove possible microorganisms and impurities, and ensuring the sterility of the culture solution;

[0034] S4: dispensing the filtered culture solution into sterile culture bottles, sterilizing the culture bottles by using a high-pressure steam sterilizer or a dry heat sterilizer, and killing residual microorganisms;

[0035] S5: adjusting the pH value of the culture solution by using an acid or alkali solution.

[0036] The additives are serum, amino acids, vitamins, and antibiotics.

[0037] The culture medium is a potato sucrose culture medium.

[0038] The filter membrane is a 0.22-micron filter membrane.

[0039] The culture bottle is 100 milliliters.

[0040] The pH value of the culture solution in S5 is 7.2.

[0041] Example Two:

[0042] A cell culture solution preparation process, comprising the following steps:

[0043] S1: selecting a basic culture medium, and adding additives to the culture medium;

[0044] S2: adding the culture medium and the additives into a preparation device, so that the culture medium and the additives are dissolved in deionized water;

[0045] S3: making the dissolved culture medium flow from the upper part to the lower part of the preparation device, filtering the prepared culture solution through a filter membrane to remove possible microorganisms and impurities, and ensuring the sterility of the culture solution;

[0046] S4: dispensing the filtered culture solution into sterile culture bottles, sterilizing the culture bottles by using a high-pressure steam sterilizer or a dry heat sterilizer, and killing residual microorganisms;

[0047] S5: adjusting the pH value of the culture solution by using acid or alkali solution.

[0048] The additive is serum, amino acid, vitamin and antibiotic.

[0049] The culture medium is Chayes culture medium.

[0050] The filter membrane is 0.45 micron filter membrane.

[0051] The culture bottle is 500 milliliter.

[0052] The pH value of the culture solution in S5 is 7.4.

[0053] As shown in Figures 8-9 this example can achieve the effect of avoiding the pollution of the culture solution.

[0054] Since the preparation device comprises a round box 101, a middle cylinder 102 is arranged at the bottom center of the round box 101, and a pressing plug 209 is pressed on the upper part of the middle cylinder 102, the culture medium and the additive can be dissolved in the deionized water in the round box 101, and after the pressing plug 209 is lifted, the culture solution can flow down from the middle cylinder 102, without manually taking out the culture solution, so as to avoid the pollution of the culture solution.

[0055] As shown in Figures 8-9 this example can achieve the effect of driving the pressing plug 209 to lift or fall.

[0056] Since a vertical shaft 202 is welded at the center of the pressing plug 209, the vertical shaft 202 is vertically and slidingly connected to an L-shaped frame 201, the vertical shaft 202 is driven to slide by a hydraulic cylinder, and the rear part of the L-shaped frame 201 is connected to the rear side of the round box 101, so that the vertical shaft 202 can drive the pressing plug 209 to lift or fall when the vertical shaft 202 vertically slides on the L-shaped frame 201.

[0057] As shown in Figures 8-9 this example can achieve the effect of conveniently disassembling the pressing plug 209 from the round box 101 as a whole.

[0058] Since the rear side of the round box 101 is connected to a rear seat 103 by a screw, the L-shaped frame 201 is inserted into the rear seat 103, a fastening screw is threadedly connected to the rear seat 103 and is pressed on the L-shaped frame 201, so that the L-shaped frame 201 is conveniently disassembled from the rear seat 103, and then the pressing plug 209 is conveniently disassembled from the round box 101 as a whole.

[0059] As shown in Figures 8-9 this example can achieve the effect of uniformly mixing the culture medium and the additive in the deionized water.

[0060] The middle part of the rotating frame 205 is provided with a gear cylinder 208 due to the middle part gap fitting of the rotating frame 205 being inserted on the vertical shaft 202, a compression spring is sleeved on the vertical shaft 202, the compression spring is arranged between the L-shaped frame 201 and the gear cylinder 208, the two ends of the rotating frame 205 are both connected with a wiping frame 206 through screws, a plurality of tabs 207 are welded on the bottom of each wiping frame 206, the L-shaped frame 201 is connected with a motor 203 through screws, a gear 204 is connected on the output shaft of the motor 203 through a key, the gear 204 is engaged with the gear cylinder 208 for transmission, the motor 203 can drive the gear 204 to rotate, the gear 204 drives the gear cylinder 208 and the rotating frame 205 to rotate around the vertical shaft 202, and then drives the plurality of tabs 207 to continuously scrape on the inner bottom surface of the circular box 101, so that the culture medium and the additive are uniformly mixed in the deionized water.

[0061] The circular box 101 is connected on the upper part of the stand 401 through screws, the rear side of the bottom box 301 is welded with a square column 403, the square column 403 is slidingly connected on the stand 401 in the front-rear direction, two circular plates 404 are welded on the square column 403, the two circular plates 404 are located on the two sides of the stand 401 respectively, and two compression springs are sleeved on the square column 403 and located on the two sides of the stand 401 respectively.

[0062] As shown in the figure, Figures 8-11 This example can realize the effect of uniformly shaking the culture solution falling in the bottom box 301.

[0063] When the culture solution is mixed, the culture solution can be discharged from the middle cylinder 102, then falls in the bottom box 301, the bottom box 301 can slide forward and backward through the square column 403, the two compression springs on the square column 403 help the square column 403 and the bottom box 301 to reset, so that the bottom box 301 shakes forward and backward and then resets, and the culture solution falling in the bottom box 301 is uniformly shaken.

[0064] The upper part of the bottom box 301 is welded with a socket 306, the rear side of the filter plate 304 is welded with a plug rod 305, the plug rod 305 is inserted on the socket 306, a fastening screw is threadedly connected on the socket 306 and presses on the plug rod 305.

[0065] As shown in the figure, Figures 10-11 This example can realize the effect of less impurities in the culture solution.

[0066] After the culture solution is discharged from the middle cylinder 102 and falls on the filter plate 304, the filter plate 304 filters the culture solution once, then the culture solution falls in the bottom box 301 for collection, and the culture solution is filtered once, so that the culture solution has less impurities.

[0067] The middle rod 303 is vertically slidingly connected to the lower center of the bottom box 301, the lower side of the bottom box 301 is annularly provided with a plurality of filter membranes 302, the middle rod 303 is slidingly driven by a hydraulic cylinder, the upper end of the middle rod 303 is welded with a circular seat 504, the upper part of the circular seat 504 is welded with a conical plate 501, the conical plate 501 is vertically slidingly connected to the inside of the bottom box 301, the circular seat 504 is provided with an L-shaped hole 503, the upper side of the conical plate 501 is provided with a blocking piece 502 which is vertically slidingly driven by a hydraulic cylinder, and the blocking piece 502 can be pressed on the upper side of the L-shaped hole 503.

[0068] As shown in Figures 10-13 , this example can remove possible microorganisms and impurities, and ensure the sterility of the culture solution.

[0069] The culture solution filtered by the filter plate 304 falls on the conical plate 501, the periphery of the conical plate 501 is high and the center is low, then the culture solution flows to the L-shaped hole 503, and the culture solution flows into the bottom box 301 from the L-shaped hole 503 for collection, then the blocking piece 502 is driven to press on the L-shaped hole 503, thereby closing the L-shaped hole 503, then the middle rod 303 is driven to move downward, thereby driving the conical plate 501 to move downward relative to the bottom box 301, thereby pressing the culture solution, so that the culture solution passes through the plurality of filter membranes 302, the prepared culture solution is filtered through the filter membranes 302, possible microorganisms and impurities are removed, and the sterility of the culture solution is ensured.

[0070] The stand 401 is vertically slidingly connected with a sliding sleeve 402, the rear part of the sliding sleeve 402 is provided with a clamping groove 405 which can be inserted on the circular sheet 404 located at the rear side.

[0071] As shown in Figure 11 , this example can lock the positions of the square column 403 and the bottom box 301.

[0072] When the sliding sleeve 402 is slid, the clamping groove 405 can be clamped on the circular sheet 404 located at the rear side, thereby locking the positions of the square column 403 and the bottom box 301. The sliding sleeve 402 is threadedly connected with a fastening screw, and the sliding sleeve 402 can be fixed by the fastening screw.

Claims

1. A process for preparing cell culture medium, characterized in that, Includes the following steps: S1: Select a basic culture medium and add additives to the culture medium; S2: Add the culture medium and additives to the preparation device so that the culture medium and additives dissolve in deionized water; S3: Allow the dissolved culture medium to flow from the top to the bottom of the preparation device, filter the prepared culture solution through a filter membrane to remove any possible microorganisms and impurities, and ensure the sterility of the culture solution; S4: Dispense the filtered culture medium into sterile culture bottles and sterilize the culture bottles using an autoclave or dry heat sterilizer to kill any remaining microorganisms. S5: Adjust the pH of the culture medium using acid or alkaline solutions.

2. The cell culture medium preparation process according to claim 1, characterized in that: The additives are serum, amino acids, vitamins, and antibiotics.

3. The cell culture medium preparation process according to claim 1, characterized in that: The culture medium is one of potato sucrose medium, bean sprout juice sucrose agar medium, or Czapek's medium.

4. The cell culture medium preparation process according to claim 1, characterized in that: The filter membrane is a 0.22-micrometer or 0.45-micrometer filter membrane.

5. The cell culture medium preparation process according to claim 1, characterized in that: The culture flask is 100 ml or 500 ml.

6. The cell culture medium preparation process according to claim 1, characterized in that: The pH of the culture medium in S5 is 7.2-7.

4.

7. The cell culture medium preparation process according to claim 1, characterized in that: The preparation device includes a round box (101), a middle cylinder (102) is provided at the bottom center of the round box (101), and a pressure plug (209) is pressed on the upper part of the middle cylinder (102).

8. The cell culture medium preparation process according to claim 7, characterized in that: The center of the pressure plug (209) is fixed with a vertical shaft (202), which is vertically slidably connected to the L-shaped frame (201). The vertical shaft (202) is driven to slide by a hydraulic cylinder. The rear part of the L-shaped frame (201) is fixed to the rear side of the round box (101).

9. The cell culture medium preparation process according to claim 8, characterized in that: A rear seat (103) is fixed to the rear side of the round box (101), and an L-shaped frame (201) is inserted into the rear seat (103). A fastening screw is threaded onto the rear seat (103), and the fastening screw presses onto the L-shaped frame (201).

10. The cell culture medium preparation process according to claim 9, characterized in that: The middle part of the rotating frame (205) is fitted onto the vertical shaft (202) with a clearance fit. A gear cylinder (208) is provided in the middle of the rotating frame (205). A compression spring is sleeved on the vertical shaft (202). The compression spring is located between the L-shaped frame (201) and the gear cylinder (208). Both ends of the rotating frame (205) are fixed with trowels (206). The bottom of each trowel (206) is fixed with multiple protrusions (207). A motor (203) is fixed on the L-shaped frame (201). A gear (204) is fixed on the output shaft of the motor (203). The gear (204) meshes with the gear cylinder (208) for transmission.

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