Jinma analysis of cell culture technology basic information - Database & Sql Blog Articles

I. Cell Culture Medium Cell culture is a technique that involves isolating cells from tissues and growing them in a controlled, sterile environment with appropriate temperature, pH, and nutrients. This process allows for the propagation and maintenance of cell populations, which are widely used in medical and genetic research. Commonly cultured cells include peripheral blood lymphocytes, skin fibroblasts, and long-term in vitro cell lines. Peripheral blood lymphocyte cultures are particularly popular due to their simplicity, quick results, and reusability, making them ideal for clinical chromosome analysis. During in vitro culture, cells may undergo spontaneous transformation or be engineered into permanent cell lines. These lines often exhibit aneuploid characteristics, though this might not always be evident in cloned cell lines. The success of cell culture depends on replicating the in vivo conditions as closely as possible. II. Environmental Conditions for Cell Culture 1. **Sterile Environment** The culture environment must be free from contaminants. Once removed from the body, cells lose their natural defenses against microbes and toxins. Contamination or accumulation of metabolic byproducts can lead to cell death. Maintaining a clean, non-toxic environment is essential for survival. 2. **Temperature Control** Human cells typically require a constant temperature of 36.5°C ± 0.5°C. Deviations beyond this range can disrupt metabolism and cause cell death. Cells are more resilient to low temperatures than high ones. At 39–40°C, some cells may recover after one hour, but at 41–42°C, most die, and at 43°C, all cells perish within an hour. 3. **Gas Environment** Oxygen and carbon dioxide are critical for cell growth. Oxygen supports energy production, while CO₂ helps maintain the pH of the medium. Most cells thrive in a 95% air and 5% CO₂ atmosphere. pH levels should remain between 7.2 and 7.4, with sodium bicarbonate (NaHCO₃) commonly used to adjust it. HEPES buffer is preferred in open systems for stable pH control. 4. **Culture Medium** The medium provides nutrients and growth factors. It can be synthetic or natural. Synthetic media contain defined components like amino acids and vitamins, while natural media, such as fetal bovine serum, provide growth factors and adhesion molecules. III. Equipment and Facilities for Cell Culture 1. **Laboratory Design** Aseptic conditions are crucial. Labs are divided into areas for preparation, operation, and cleaning. Clean benches, incubators, centrifuges, and microscopes are standard equipment. 2. **Common Tools** Glassware, flasks, Petri dishes, pipettes, and centrifuge tubes are essential. Each has specific uses depending on the experiment. For example, flasks are used for cell attachment, while Petri dishes are for open cultures. 3. **Cell Morphology** Cells can grow as adherent or suspension types. Adherent cells attach to surfaces, like fibroblasts and epithelial cells, while suspension cells float freely in the medium. IV. Analysis and Maintenance of Cultured Cells Cultured cells vary in shape based on the support they grow on. Healthy cells appear transparent under a microscope, while stressed or damaged cells show irregularities, such as increased contrast or cytoplasmic granules. V. Cleaning and Disinfection Proper cleaning of glassware, rubber stoppers, and plastic items is vital to prevent contamination. Methods include soaking, brushing, pickling with acid solutions, and rinsing. Disinfection techniques like UV light, steam sterilization, and chemical agents ensure a sterile environment. VI. Importance of Sterility Contamination is the biggest threat in cell culture. Strict adherence to protocols, including regular disinfection and aseptic handling, is necessary to avoid failure. Antimicrobial agents are also used to protect cultures from bacterial or viral threats. By maintaining optimal conditions and following rigorous procedures, researchers can successfully cultivate and study cells for various scientific applications.

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