Cell Culture Medium Optimization- The primary aim of maintaining cell lines in high standard is to maximize the reproducibility and reliability of the experiment. The Cell culture is the process by which cell are grown under the controlled environment with appropriate nutrition e.g. cell culture medium, essential amino acid and Protein source (Difference Between Primary And Secondary Cell Lines).
Once cell are isolated form it’s targeted tissue, cells are subsequently maintained under the controlled environment 5% CO2 at 37°C with >90% humidity supplement. Conditions may vary from cells to cells, but most of the cells can be grown in same kind of controlled environment. Key elements of in vitro cell cultures are culture medium, growth hormone supplements, protein source and controlled environment.
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Composition of Cell Culture Media and Function
Cell culture media is composed of a variety of different nutrients and supplements that are essential for the growth and survival of cells in vitro (MTT Test). The most common cell culture media components include:
- A source of energy (usually glucose or another sugar)
- Amino acids
- Inorganic salts
- Vitamins
- Trace elements
- Growth factors
- Antibiotics (optional)
Cell culture media is composed of many different types of molecules including proteins, carbohydrates, lipids, and nucleic acids. Each type of molecule has a specific function in supporting cell growth and development.
Proteins provide the main source of nitrogen and essential amino acids needed for cell growth. Carbohydrates are important for energy production and cellular metabolism. Lipids are necessary for cell membrane structure and function. Nucleic acids are required for DNA replication, transcription, and translation.
| Sn. | Component | Function |
| 1 | Medium (MEM, RPMI, DMEM) | Source of Energy and Growth hormone |
| 2 | Essential Amino acid | Building protein blocks |
| 3 | Glucose | Energy Source |
| 4 | Vitamin | Cell survival and growth |
| 5 | Phenol Red | pH Indicator |
| 6 | Bicarbonate /HEPES buffer | Balancing pH in the media |
| 7 | Temperature (37°C) | Physiologic Temperature |
| 8 | 5% CO2 | To maintain pH |
| 9 | Humidity (90%) | To provide oxygen |
Different types of Culture Medium and their Uses
| Sn. | Media Type | Function |
| 1 | MEM | Primary and diploid culture |
| 2 | DMEM | Modification of MEM containing increased Level of amino acids and vitamins. Supports a wide range of cell types including hybridomas |
| 3 | GMEM | Glasgow’s modified MEM was defined for BHK-21 cells |
| 4 | RPMI 1640 | Originally derived for human leukemic cells. It supports a wide range of mammalian cells including hybridomas |
| 5 | Iscoves DMEM | Further enriched modification of DMEM which supports high density growth |
| 6 | Leibovitz L-15 | Designed for CO2 free environments |
Those above mentioned factors play important role in the growth of the cell lines. Different kinds of mediums are available to maintain the cell lines e.g. DMEM, MEM, RPMI 1640. Medium captains growth hormones and essential nutrition for growth and differentiation of the cell lines. Other growth factors such as proteins and amino acid derive from fetal bovine serum (FBS), bovine calf serum, equine serum, and porcine serum.
Cell Culture Medium Optimization Formulae
To prepare 10% Complete Medium
For 100mL
- 10% FBS- 10 mL
- Medium – 88 mL
- 1% L- Glutamine – 1mL
- 1% Antibiotic – 1mL
pH could be maintained by adding the buffer solution and 5% Co2.
Growth of the cell lines should be checked twice in a day to observe the pH and if it has become pale yellow in color, medium should be changed immediately.
Cell Culture Medium Optimization Requirement
In order to grow and proliferate, mammalian cells require a nutrient-rich environment. This environment is typically provided in the form of a liquid cell culture medium. The composition of the cell culture medium can have a significant impact on the proliferation and function of the cells, making it critical to optimize the medium for each specific application in Genetic Toxicology.
There are many different types of cell culture media available, each with its own advantages and disadvantages. For example, some media are more expensive than others, while some are more complex to prepare. In addition, certain media formulations may be more suitable for certain applications than others.