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

I. Cell Culture Medium Cell culture is a technique that involves removing cells from body tissues and growing them in a controlled, sterile environment with appropriate temperature, pH, and nutrients. This process allows for the propagation and maintenance of single or multiple cell populations. The most commonly used cell types in medical genetics research include peripheral blood lymphocytes, skin fibroblasts, and long-term in vitro cell lines. Peripheral blood lymphocyte cultures are widely used due to their short time frame, simple procedures, and reusable materials, making them ideal for clinical chromosome analysis. In vitro cultured cell lines may undergo spontaneous transformation or be engineered into permanent lines, which can be passaged indefinitely. These permanent lines often exhibit aneuploid characteristics, though this may not always be visible in cloned lines. II. Environmental Conditions for Cell Culture 1. Non-Polluting Environment The culture environment must be free from toxins and microbes. Once removed from the body, cells lose their natural defenses, making contamination or metabolic buildup dangerous. Maintaining a clean environment and regularly removing waste products are essential for cell survival. 2. Constant Temperature Human cell cultures require a stable temperature of around 36.5°C ± 0.5°C. Deviations can disrupt metabolism or cause cell death. Cells tolerate low temperatures better than high ones. Prolonged exposure to temperatures above 40°C can lead to irreversible damage. 3. Gas Environment Oxygen and carbon dioxide are critical for cell survival. Oxygen supports energy production, while CO₂ helps maintain the medium’s pH. Most cells thrive in a pH range of 7.2–7.4. Sodium bicarbonate (NaHCO₃) is commonly used to buffer pH, while HEPES provides stability in open systems. 4. Culture Medium The medium supplies nutrients and supports cell growth. It can be synthetic or natural, with serum being the most common natural source. Synthetic media contain specific components like carbohydrates, amino acids, and growth factors. III. Facilities and Equipment for Cell Culture 1. Laboratory Design A sterile environment is crucial. Workspaces should minimize microbial exposure, with designated areas for aseptic operations, storage, and cleaning. Proper airflow and cleanliness are essential for maintaining sterility. 2. Common Equipment Key tools include laminar flow hoods, incubators, centrifuges, microscopes, and sterilization units. Each plays a role in maintaining aseptic conditions and supporting cell growth. 3. Culture Vessels Glassware such as flasks, petri dishes, and tubes are essential. They must be transparent, non-toxic, and durable. Reusable glassware requires thorough cleaning and sterilization before use. 4. Cell Morphology Cells can grow as adherent or suspension types. Adherent cells attach to surfaces, while suspension cells float in the medium. Different cell types, such as fibroblasts, epithelial cells, and migratory cells, have distinct morphological features. IV. Analysis and Maintenance of Cultured Cells Cultured cells vary in shape depending on their attachment. Healthy cells appear uniform under a microscope, with clear nuclei and cytoplasm. Changes in morphology, such as increased contrast or granules, may indicate poor health or metabolic issues. V. Cleaning and Disinfection Proper cleaning of glassware, rubber stoppers, and plastic items is vital. Methods include soaking, brushing, pickling with acid solutions, and rinsing. Disinfection techniques like UV light, steam sterilization, and chemical agents help prevent contamination. VI. Contamination Prevention Contamination is a major risk in cell culture. Strict adherence to protocols, including regular disinfection and aseptic techniques, is necessary to ensure successful experiments. Each step must be carefully followed to avoid failure. This detailed guide ensures that all aspects of cell culture are understood and properly managed, promoting reliable and reproducible results.

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