Cell Culture Freezing Medium: The Complete FAQ Guide In 2025

1.What idea do you get from cell culture freezing medium?

Cell Culture Freezing Medium-Picture Courtesy: Vitechltd

Have you thought how different cells are available for experimentation and manufacturing, despite of being so vulnerable? Like how they endure the environment to remain functional and useful? Cell culture freezing medium is responsible behind the effectiveness of cell lines.

You can define cell culture freezing medium as a unique liquid prepared to preserve different cells at extremely low temperatures. This medium is comprised of some valuable components that keep different cells protected and working even after storing for a long time.

2.Do you know the advantages of cell culture freezing medium?

Cell culture freezing medium secures an important place in the field of research and pharma production, but do you know its advantageous role? This topic is crafted to manifest you the advantages you can get from this useful product.

Encouraging the cell banks:

Encouraging The Cell Banks-Picture Courtesy: JOINN Biologics

Cells are used in different experiments and they are also required for further research but how to get them in viable form? Cell culture freezing medium encourages the cell banks that is an authentic source to procure these cell lines and you can get different kinds of cells for carrying out experiment.

Keeping the cell structure sturdy:

Keeping The Cell Structure Sturdy-Picture Courtesy: Texcell

Cell culture freezing medium ensures the complete safety of cell lines. You can observe that the structure of these cells remain sturdy and no damage happens due to freezing or thawing. This medium contains ingredients that support the preservation of all cell components.

Protecting the vitality of cells:

Protecting The Vitality Of Cells-Picture Courtesy: Precision Antibody

Cells lines remain effective and working even after storing in cell culture freezing medium. It proves that this freezing medium has great tendency to protect the vitality of cells. Cryoprotective agent is present in medium that prevents destruction of cell lines during freezing and thawing and you attain a viable form of different cells.

Supporting the prolonged cell storage:

Supporting The Prolonged Cell Storage-Picture Courtesy: 53Biologics

Cell culture freezing medium supports the prolonged cell storage by the help of cryopreservation? Do you know what cryopreservation is? This is the process of protecting cells during freezing and thawing by using cryoprotectants, which avoid crystal formation in cells that can lead to cell damage.

You can store cells for longer time without fearing about any impairment and utilize your required cells when it is needed either in experiment or production.

Offering experimental feasibility:

Offering Experimental Feasibility-Picture Courtesy: News-Medical.net

Research can be so diversified and you can need any cell line anytime. Preservation of these cell lines by the help of cell culture freezing medium provides you convenience in obtaining cell of your choice upon demand. This saves your time and resources and offers experimental feasibility.

Proving cost efficient:

Proving Cost Efficient-Picture Courtesy: Drug Target Review

After investing in cell culture freezing medium, you can find it cost efficient. You don’t need to nourish cell lines every time as you have the facility to store different cells that can be used instantly.

Aiding in shipping of cell lines:

Aiding In Shipping Of Cell Lines-Picture Courtesy: MedicaleExpo e-Magazine

When you get a reliable source of protecting cell lines then shipping becomes easier. The requirement of cells in remote areas can be fulfilled by utilizing cell culture freezing medium that transports the cells safely.

Facilitating cell therapy:

Facilitating Cell Therapy-Picture Courtesy: Atlantis Bioscience

You are witnessing the advancement in field of medicine and cell therapy is becoming common. The demanded cells can be taken from the storage of these cells. Cell culture freezing medium provides an easy way to obtain cells upon demand.

Minimizing contamination:

Minimizing Contamination-Picture Courtesy: uBriGene Biosciences

Do you sense that liquid state promotes microbial growth? Yes, this is right and that’s the reason behind preserving the cells in cell culture freezing medium. Even after their storage in this medium, these cells are shifted to nitrogen liquid so that their integrity does not affect during thawing. You can decrease the chances of contamination and get functional cells by this freezing medium.

3.What can be the applications of cell culture freezing medium?

This topic educates you about the sectors where cell culture freezing medium contributes as an impactful factor.

Applications of cell culture freezing medium:	Image:
Manufacturing of biopharmaceuticals: Do you know the biopharmaceuticals that require cell lines? Vaccines, hormones, antibodies and enzymes need different cells for their production. Cell culture freezing medium facilitates the availability of desirable cell lines for such important products that endorses the importance of cell culture freezing medium in pharmaceutical industry.	Picture Courtesy: Science
Research & development: Are you aware of the fact that research demands uniform supply of cell lines to carryout different experiments either you have to discover a drug or to know about toxicity of it. Assays are carried out to get final result and assays are based on cell lines. Cell culture freezing medium offers you a continuous supply of viable cells that facilitates your research work smoothly.	Picture Courtesy: Drug Target Review
Cell storage: Do you realize that cell culture freezing medium supports the cell stocks? Yes, this is true because the cryopreservative agent in this medium functions to protect the cell lines from being impaired and also freezing medium avoid microbial presence. Cell lines storage has become convenient due to cell culture freezing medium.	Picture Courtesy: Eurofins USA

4.What packaging containers are chosen to dispense cell culture freezing medium?

Are you known to the value of packaging container? It provides protection to the formulation during storage and it has to be appropriate according to formulation’s needs. Here you get to learn about the packaging containers that are chosen to dispense cell culture freezing medium.

Packaging container	Image:
Plastic bottles: You can observe that the cell culture freezing medium is available in bioprocessing containers. These containers are look like bottle and covered with plastic lid to keep the container airtight. These bottles contain bulk amount of formulations like 50ml. The culture freezing medium present in these containers can be used only for once. Such containers are used for large scale research or manufacturing of biopharmaceuticals.	Picture Courtesy: Science Cell
Cryopreservation vials: These vials are different from commonly known vials that are cylindrical in shape and sealed by aluminum caps. Cryopreservation vials are plastic tube like container that is closed by the plastic screw cap to keep formulation leakage-free. The purpose of this small container is storage of cell lines and these vials are placed in a specialized container that possesses controlled freezing temperature. These vials contain small amount of cell culture freezing medium like 2ml. These containers have great tendency to store cell lines in cell culture freezing medium in extremely low temperature and that’s the reason these are highly preferable.	Picture Courtesy: RoosterBio

5.Can you explain the storage protocol of cell culture freezing medium?

You must be very concerned about the storage protocols of cell culture freezing medium. Any negligence can cause you a big trouble in future. Here you get to know the important aspects of storage protocol that you have to look after to keep the cell culture freezing medium stable.

Freezing containers:

Freezing Container-Picture Courtesy: Carl ROTH

You must know about the freezing container that is appropriate to maintain the extremely low temperature. It is particularly designed to maintain cooling rate of cell lines preserved in cell culture freezing medium. This container acquires a medium that is usually isopropyl alcohol that keeps the temperature controlled. Cryopreservation vials are placed in this container for proper storage. Mechanical freezer is required to place this container in it and vials get the desirable temperature due to mechanism this freezing container follows.

Temperature:

Freezing Temperature Is Required To Store Cell Culture Freezing Medium-Picture Courtesy: Technology Networks

Do you know the suitable temperature for the storage of cell culture freezing medium? You have learnt that this medium demands for frozen temperature, -20 ℃ is considered as the ideal temperature for storing cell culture freezing medium.

When you have needed to use this medium, you can thaw this medium and store it on 2-8 ℃ but it can only be used for very few days.

Expiry date:

Expiry date

Like other products, cell culture freezing medium can also get expired. This information is very critical as storage temperature has huge influence on expiration of this medium.

When cell culture freezing medium is stored in frozen temperature like -20 ̊℃ to -5 ℃, it can be used for 1 year after its production.

Do you know the meaning of thawing? It is the process of changing the frozen solid state into liquid state. Thawing is done to cell culture freezing medium by storing it on 2-8 ̊℃. After that you can use it within 5 days. You must not place it freezer after thawing it.

6.What components are used to make cell culture freezing medium?

Are you inquisitive to know about the components of cell culture freezing medium? Fundamentally this medium is made from three important components which are discussed below:

Components of cell culture freezing medium	Image
Cryoprotective agent: Have you thought which chemical is preventing the frozen state of cells so that they don’t lose their viability? It is Cryoprotective agent. It is added in cell culture freezing medium so that it can prevent crystal formation and cell components do not get destabilized by getting frozen. Examples include dimethyl sulfoxide (DMSO), glycerol.
Basal medium: Do you wonder how cells get basic nutrients while being preserved in cell culture freezing medium? Basal medium has it all. This medium contains all the nutrients a cell needs to survive like amino acids and vitamins. Examples include Dulbecco modified eagle medium (DMEM), Roswell park memorial institute (RPMI 1640)
Serum: Do you know cells need support during the freezing and thawing? For this purpose serum is added into the cell culture freezing medium. It provides protein and other important nutrients that can help cells to remain functional and effective. Example includes fetal bovine serum.

7.Can you define the production of cell culture freezing medium on industrial basis?

Are you agreed with the importance of cell culture freezing medium? This medium has great demand whether you want to perform an experiment or intend to produce any pharmaceutical product. Production of cell culture freezing medium requires controlled environment and efficacious equipment to get the desirable product. Sterile conditions are maintained throughout the process to prevent the sources of contamination.

 

Production of cell culture freezing medium	Information:
Preparation of components: Picture Courtesy: GetReskilled	a. Cryoprotective agent: You want cryoprotective agent to produce the cell culture freezing medium, for this purpose you prepare cryoprotective agent first. At first, you obtain phosphate buffer by dissolving suitable phosphate salts in water. As you prepare the phosphate buffer, you add cryoprotectant like dimethyl sulfoxide in it and stir it gently to get the solution form. Now you can filter it by using sterile filters. Your cyroprotective agent is ready to incorporate for production of cell culture freezing medium. b. Basal medium: You require basal medium, so to prepare it you need powdered form of DMEM, water and sodium bicarbonate. Initially you dissolve DMEM in water and stir it until it gets completely dissolved. After that you add sodium bicarbonate in DMEM solution and stir it well. You can add HCL or NaOH to attain a desirable pH of solution. Now you perform filtration by using sterile filters. c. Serum: To make serum like bovine fetus serum, you have to collect blood from bovine fetuses. After sourcing the blood sample, you put it for clotting by natural process. After clotting, you utilize centrifugation method to isolate serum from blood. Now you filter the serum by using sterile filters. Serum is ready to be used for production of cell culture freezing medium. You need to assess the quality of above components before including it into production process.
Mixing of components: Picture Courtesy: European Pharmaceutical Review	You need to follow the process of mixing of all components to get cell culture freezing medium. You must remember that the manufacturing area must have aseptic conditions and temperature and humidity must keep in limit. First you take a precise amount of each component and mix them together by using a homogenizer. You need to understand that the final product must have a uniform mixture.
Filtration:	After you got a homogenized mixture, you have to filtrate it from impurities to acquire a sterile mixture.
Filling and sealing: Picture Courtesy: Packaging Strategies	Cell culture freezing medium is regarded as sterile product and you have to utilize aseptic filling line to fill and seal it in desirable packaging container. Filling must be accurate and whole process must be carried out in closed environment to refrain contaminants inclusion in final product. Vial filling line is preferred for filling and sealing the cell culture freezing medium in cryovials. For filling cell culture freezing medium in bottles, you can chose plastic bottle filling machine. Process includes the washing of containers with sterile water, and then sterilization is carried out by means of heat or ultrasonic waves. After sterilization of containers, they are moved to filling unit. After filling, these containers are closed with plastic caps.
Final sterilization: Picture Courtesy: Zealway	For final sterilization, you must chose autoclaves that are specific for heat-sensitive products.
Quality control:	You can check the quality of cell culture freezing medium by performing various tests like sterility test, pH test, osmolality test etc.
Labeling:	After confirming the quality, now you can label the formulation with necessary information.
Documentation: Picture Courtesy: Knowmax	You must document all valuable information related to production of cell culture freezing medium.
Storage: Picture Courtesy: Presto Testing Instruments	You store the cell culture freezing medium on very low temperature to keep it effective for use.

8.What equipment is required for production of cell culture freezing medium?

Production of cell culture freezing medium requires well organized and efficient equipment to perform different steps. All equipment must follow the mandatory standards and must have the capability to maintain sterile conditions during process.

Equipment	Image
Water treatment system: AIPAK Water Treatment System	Water which you use during the production of cell culture freezing medium is sourced by treating this water from water treatment system. This system has capability to provide high purity water.
Mixing vessel:	Mixing vessel is helpful in preparation of components, which are used in production of cell culture freezing medium.
Centrifuge:	You can use centrifuge to separate serum from blood sample.
Homogenizer: AIPAK Homogenizer	Homogenizer is specifically designed equipment that facilitates the even mixing of components of cell culture freezing medium. High shear mixing is behind the homogenization of the mixture.
Sterile filters:	Sterile filters contain micro membrane that retains impurities and microorganism and you get a sterile formulation after passing the mixture from it.
Plastic bottle filling machine: Picture Courtesy: Rotzinger	You can use plastic bottle filling machine to fill cell culture freezing medium in bottles. It is needed to wash and sterilize the bottles before filling the medium in it. This ensures the sterility of product. You can fill precise amount of product in bottles and then covered it in capping unit by plastic caps.
Vial filling line: AIPAK Vial Filling Machine	Vial filling line is chosen on the basis of its ability to maintain aseptic conditions during whole procedure. It has washing unit, sterilization unit, filling and sealing units. Moreover you can see, it is protected by restricted access barrier system.
Autoclaves: AIPAK Autoclave	You consume autoclave to sterilize the final product for the last time, after that product is subjected for labeling and distribution. This autoclave is particularly designed to sterilize the heat sensitive products and cell culture freezing medium contains serum which is heat-sensitive.
Labeling machine: AIPAK Labeling Machine	Labeling machine helps you to label the cell culture freezing medium with correct information. You can apply labels accurately by adhesive method.
Freezers:	Freezers are supposed to be important equipment for production of cell culture freezing unit as the components or the final product need very low temperature for storage. This keeps the product stable and effective.

9.What aspects are important for cell culture freezing medium?

Important Aspects Of Cell Culture Freezing Medium-Picture Courtesy: Thermos Fisher Scientific

Do you know the important aspects of cell culture freezing medium? This topic presents you brief understanding of the things that have great influence on the purity and stability of cell culture freezing medium so that you focus on them.

Important aspects	Information
Regulatory compliance:	You must make sure that the whole process must be competent to guidelines proposed by good manufacturing practices (GMP). This feature helps you to attain a high quality product and you don’t need to be worried about its quality. You must ensure that machines that are taking part in production of cell culture freezing medium must be complained to regulatory standards. Keep this in your mind that documentation is as necessary as any other step. It must document important information according to regulatory standards.
Composition of medium:	You must decide the composition of freezing medium according to cell lines type you want to preserve. Do you know the standard medium composition for research purpose? It contains 90% FBS /10% DMSO, but for clinical purpose you can prefer serum free cell culture freezing medium. Cryoprotectants must be added in suitable amount so that it only shows profound effects, not become a source of toxicity.
Temperature and storage:	You must remember that cell culture freezing medium must remain stable in extremely low temperature like up to -20 ̊℃. When you preserve cells in it, then there freezing rate must be gradual like -1 ̊℃ per min that reaches to -80 ̊℃ to -70 ̊℃. For long term storage, you must store cell culture freezing medium on vapors phase of nitrogen liquid. This keeps it safe and viable.

10.Can you narrate the expected challenges of production of cell culture freezing medium? How can you solve it?

Challenges Of Cell Culture Freezing Medium Production-Picture Courtesy: ATUM

Challenges	How to solve?
Cryoprotectant: You may experience this problem that crystal formation occurs due to inappropriate amount of cryoprotectants, which leads to huge loss. This problem affects the cell components and it gets damage.	You must add precise amount of suitable cryoprotectant and must be very careful about it because this costs you so much trouble.
Freezing rates: Sometimes cell culture freezing medium does not get stored in optimal freezing rates, which causes crystal formation. This causes damage to cell.	You must use freezers that can maintain an optimal freezing rate for the storage of cell culture freezing medium.
Contamination: Although you try to keep aseptic conditions throughout the process but sometimes cell culture freezing medium gets contaminated.	You need to follow guidelines of good manufacturing practices to refrain from issues of contamination. You must utilize all means that can contribute in maintaining the aseptic environment.

11.What precautionary measures are integral for handling cell culture freezing medium?

Precautionary Measures For Cell Culture Freezing Medium-Picture Courtesy: Charles River Laboratories

You are working in a pharmaceutical industry or you are carrying out a research, you must be well-informed about the precautionary measures that are integral for handling cell culture freezing medium.

You must know it well that cell culture freezing medium contains components that can be hazardous for you, like DMSO. It can be carcinogenic. You must have to wear gloves, lab coat, and face shield to deal with this freezing medium.

As you learnt it before that we prefer nitrogen liquid for storing cell culture freezing medium for long time. So keep this in your mind that nitrogen liquid is severely cold that it can cause you frost bites. When you are placing this medium in nitrogen liquid, you must have to wear heat-insulated gloves, gown and proper face shield to protect yourself from damage.

You must look for proper storage conditions for keeping cell culture freezing medium. This requires lowest temperature like -20 ̊℃.

For avoiding the risk of contamination, you must use laminar flow hood to perform handling of cell culture freezing medium.

You must know the appropriate composition of cell culture freezing medium according to types of cell. This helps you in placing the cell lines in suitable cell culture freezing medium for preservation.

You must know the expiry of cell culture freezing medium, as you can use this within one year after production when stored on -20 ̊℃,but as you place it for thawing, after that you have to use it within 5 days.

Conclusion:

Cell culture freezing medium contributes in manufacturing of biopharmaceutical and research. It demands for GMP compliant procedures and machines. Storage protocols also need to be maintained to attain a high quality product. You must be careful about handling it. If you are willing to get more information, you can contact AIPAK ENGINEERING at any time!