Trang 1 ANIMAL STEM CELL CULTUREPHAN LU CHINH NHAN Trang 2 NUÔI CẤY TẾ BÀOĐỘNG VẬT LÀ GÌ?¡Animal Cell culture Trang 3 TẠI SAO PHẢI NUÔI CẤY TẾ BÀO ĐỘNGVẬT?Areas where cell culture tech
ANIMAL STEM CELL CULTURE PHAN LU CHINH NHAN STEM CELL INSTITUTE OCT 2021 NUÔI CẤY TẾ BÀO ĐỘNG VẬT LÀ GÌ? ¡ Animal Cell culture ¡ Animal Cell cultivation TẠI SAO PHẢI NUÔI CẤY TẾ BÀO ĐỘNG VẬT? Areas where cell culture technology is currently playing a major role ¡ Model systems for Studying basic cell biology, interactions between disease causing agents and cells, effects of drugs on cells, process and triggering of aging & nutritional studies ¡ Toxicity testing Study the effects of new drugs ¡ Cancer research Study the function of various chemicals, virus & radiation to convert normal cultured cells to cancerous cells TẠI SAO PHẢI NUÔI CẤY TẾ BÀO ĐỘNG VẬT? ¡ Virology Cultivation of virus for vaccine production, also used to study there infectious cycle ¡ Genetic Engineering Production of commercial proteins, large scale production of viruses for use in vaccine production e.g polio, rabies, chicken pox, hepatitis B & measles ¡ Gene therapy Cells having a functional gene can be replaced to cells which are having non-functional gene ƯU NHƯỢC ĐIỂM NUÔI CẤY TẾ BÀO ĐỘNG VẬT? Advantages of Animal Cell Culture: ¡ a Controlled physiochemical environment (pH, temperature, osmotic pressure, O2, etc.) ¡ b Controlled and defined physiological conditions ¡ c Homogeneity of cell types (achieved through serial passages) ¡ d Economical, since smaller quantities of reagents are needed than in vivo ¡ e Legal, moral and ethical questions of animal experimentation are avoided ƯU NHƯỢC ĐIỂM NUÔI CẤY TẾ BÀO ĐỘNG VẬT? Disadvantages of Animal Cell Culture: ¡ a Expertise is needed, so that behaviour of cells in culture can be interpreted and regulated ¡ b Ten times more expensive for same quantity of animal tissue; therefore, reasons for its use should be compelling ¡ c Unstable aneuploid chromosome constitution a Sterile Work Area: CÁC ĐIỀU KIỆN CẦN – ĐỦ CHO NUÔI CẤY TẾ BÀO ĐỘNG VẬT ¡ Where possible, a separate room should be made available for clean cell culture work This room should be free of “through traffic” and, if possible, equipped with an air flow cabinet which supplies filtered air around the work surface ¡ A HEPA (High Efficiency Particle Air Filter) filtered air supply is desirable but not always affordable Primary animal tissue and micro-organisms must not be cultured in or near the cell culture laboratory and the laboratory must be specifically designated for clean cell culture work CÁC ĐIỀU KIỆN CẦN – ĐỦ CHO NUÔI CẤY TẾ BÀO ĐỘNG VẬT b Incubation Facilities: ¡ In addition to an airflow cabinet and benching which can be easily cleaned, the cell culture laboratory will need to be furnished with an incubator or hot room to maintain the cells at 3040°C The incubation temperature will depend on the type of cells being cultivated Insect cells will grow best at around 30°C while mammalian cells require a temperature of 37°C CÁC ĐIỀU KIỆN CẦN – ĐỦ CHO NUÔI CẤY TẾ BÀO ĐỘNG VẬT c Refrigerators and Freezer: ¡ Both items are very important for storage of liquid media at 4°C and for enzymes (e.g., trypsin) and some media components (e.g., glutamine and serum) at -20°C A refrigerator or cold room is required to store medium and buffers ¡ A freezer will be needed for keeping pre aliquot stocks of serum, nutrients and antibiotics Reagents may be stored at a temperature of 20°C but if cells are to be preserved it may be necessary to provide liquid nitrogen or a -70°C freezer CÁC ĐIỀU KIỆN CẦN – ĐỦ CHO NUÔI CẤY TẾ BÀO ĐỘNG VẬT d Microscopes: ¡ A simple inverted microscope is essential so that cultures can be examined in flasks and dishes It is vital to be able to recognize morphological changes in cultures since these may be the first indication of deterioration of a culture ¡ A very simple light microscope with x 100 magnification will suffice for routine cell counts in a haemocytometer, although a microscope of much better quality will be required for chromosome analysis or autoradiography work CÁC ĐIỀU KIỆN CẦN – ĐỦ CHO NUÔI CẤY TẾ BÀO ĐỘNG VẬT (b) Synthetic Media: ¡ Synthetic media are prepared artificially by adding several organic and inorganic nutrients, vitamins, salts, serum proteins, carbohydrates, cofactors, etc Different types of synthetic media can be prepared for a variety of cells and tissues to be cultured Synthetic Media are of Two Types: ¡ Serum containing media (media containing serum) ¡ Serum- free media (media without serum)