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细胞培养环境
One of the major advantages of cell culture is the ability to manipulate the cell culture environment—the physicochemical (i.e., temperature, pH, osmotic pressure, O2 and CO2 tension) and the physiological environment (i.e., hormone and nutrient concentrations) in which the cells propagate.
A well-regulated cell culture environment is important for the health, growth, and productivity of your cells. Find comprehensive information on essential environmental parameters such as temperature, humidity, gas composition, and culture media. Read below about ways to optimize your cell culture conditions and enhance the quality and reproducibility of your experimental results.
Note that cell culture conditions vary for each cell type.
The consequences of deviating from the culture conditions required for a particular cell type can range from the expression of aberrant phenotypes to a complete failure of the cell culture. We recommend that you familiarize yourself with your cell line of interest and closely follow the instructions provided with each product you are using in your experiments.
Cell Culture pH levels
Most normal mammalian cell lines grow well at pH 7.4, and there is very little variability among different cell lines. Some transformed cell lines have shown to grow better at slightly more acidic environments (pH 7.0–7.4), and some normal fibroblast cell lines prefer slightly more basic environments (pH 7.4–7.7).
Insect cells are cultured in growth media that are usually more acidic than those used for mammalian cells. Insect cell lines such as Sf9 and Sf21 grow optimally at pH 6.2. Unlike mammalian cell cultures, the pH rises gradually as the insect cells grow, but usually does not exceed pH 6.4. Eventually though, as higher cell densities are reached, both insect and mammalian cell growth media will experience a reduction in pH.
Cell Culture CO2 levels
While most researchers usually use 5–7% CO2 in air, 4–10% CO2 is common for most mammalian cell culture experiments. The growth medium affects the pH of the cell culture environment and buffers the cells in culture against changes in the pH. Usually, this buffering is achieved by including an organic (e.g., HEPES) or CO2-bicarbonate based buffer. Because the pH of the medium is dependent on the delicate balance of dissolved carbon dioxide (CO2) and bicarbonate (HCO3–), changes in the atmospheric CO2 can alter the pH of the medium. Therefore, it is necessary to use exogenous CO2 when using media buffered with a CO2-bicarbonate based buffer, especially if the cells are cultured in open dishes or transformed cell lines are cultured at high concentrations.
Each medium has a recommended CO2 tension and bicarbonate concentration to achieve the correct pH and osmolality; refer to the media manufacturer’s instructions for more information.
Optimal temperatures for various cell lines
The optimal temperature for cell culture largely depends on the body temperature of the host from which the cells were isolated, and to a lesser degree on the anatomical variation in temperature (e.g., temperature of the skin may be lower than the temperature of skeletal muscle). Overheating is a more serious problem than underheating for cell cultures; therefore, the temperature in the incubator is often set slightly lower than the optimal temperature.
- Most human and mammalian cell lines are maintained at 36°C to 37°C for optimal growth.
- Insect cells are cultured at 27°C for optimal growth; they grow more slowly at lower temperatures and at temperatures between 27°C and 30°C. Above 30°C, the viability of insect cells decreases, and the cells do not recover even after they are returned to 27°C.
- Avian cell lines require 38.5°C for maximum growth. Although these cells can also be maintained at 37°C, they will grow more slowly.
- Cell lines derived from cold-blooded animals (e.g., amphibians, cold-water fish) tolerate a wide temperature range between 15°C and 26°C.
仅供科研使用，不可用于诊断目的。