cellculture biochemp
اسلاید 1: 1
اسلاید 2: Introduction Cell culture is the process by which prokaryotic, eukaryotic or plant cells are grown under controlled conditions. But in practice it refers to the culturing of cells derived from animal cells.2
اسلاید 3: Basic factor in cell culture technologyFirst development was the use of antibiotics which inhibits the growth of contaminants.Second was the use of trypsin to remove adherent cells to subculture further from the culture vesselThird was the use of chemically defined culture medium.3
اسلاید 4: Cell culture Terminology Tissue culture: In vitro cultivation of organs, tissues & cells at defined temperature using an incubator & supplemented with a medium containing cell nutrients & growth factors is collectively known as tissue cultureOrgan culture:A three dimensional culture of un disaggregated tissue retaining some or all of the features of the tissue in vivo Primary explants:Derived from an explant, directly from the animal Usually only survive for a finite period of time Involves enzymatic and/or mechanical disruption of the tissue and some selection steps to isolate the cells of interest from a heterogeneous population (frog /tissue regeneration) chicken in egg / transgenic mics /tumor cellDifferent types of cell grown in culture includes connective tissue elements such as fibroblasts, skeletal tissue, cardiac, epithelial tissue (liver, breast, skin, kidney) and many different types of tumor cells.Histotypic culture 4
اسلاید 5: TerminologyClone :A population derived from a single cellSub-culture :Transplantation of cells from one vessel to anotherEstablished or Continuous Cell Lines :A primary culture that has become immortal due to some transformationMost commonly tumour derived, or transformed with a virus such as Epstein-BarrOne of the most commonly used cells are Chinese Hamster Ovary cells (CHO)The SH-SY-5Y cells a human neuroblastoma derived cell linePassage Number :Number of successive sub-cultures from primary culture5
اسلاید 6: 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 studiesToxicity testing : Study the effects of new drugs Cancer research : Study the function of various chemicals, virus & radiation to convert normal culturedVirology : 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 gene6
اسلاید 7: محیط کار استریل برای انجام کارهای مربوط به کشت سلول باید تا جایی که امکان دارد یک اتاق جداگانه در نظر گرفته شود. این اتاق باید عاری از هر گونه شلوغی بوده و در صورت امکان از یک تهویه هوای فیلتر شده در محیط کار برخوردار باشد. یک فیلتر هوای (HEPA :High Efficiency Particle Air Filter ) می تواند چنین محیطی را فراهم آورد. بافت اولیه حیوانی و میکروارگانیسم ها نباید در نزدیکی آزمایشگاه کشت سلول کشت شوند و باید یک آزمایشگاه جداگانه و اختصاصی برای کار کشت سلول به صورت استریل طراحی شود. پوشش های مخصوص آزمایشگاه کشت سلول باید در ورودی آزمایشگاه پوشیده شده و لباسی که از این محیط خارج می گردد نباید مجدداً وارد آزمایشگاه شود. در صورت نیاز به استریلیتی شدید یک هود لامینار جهت حفظ بهتر از شرایط استریل پیشنهاد می گردد. در صورت استفاده از یک ماده شیمیایی خطرناک باید از هود های ورتیکال یا عمودی استفاده گردد. تمام سطوح کار، میز ها و قفسه ها و هود لامینار باید به طور مرتب با اتانول 70% یا یک پاک کننده جایگزین به درستی پاکیزه و استریل گردند. در صورتی که هود لامینار یا فیلتر های هوا در محیط کار وجود نداشت، کار کشت می تواند بر روی یک میز تمیز با کمک شعله آتش جهت استیل نمودن سطح کار و هوای اطراف آن انجام گردد. 7
اسلاید 8: Design and equipment for cell culture laboratorieslaboratory design: ideally divided into two areas:an working area reserved for handling of new material (quarantine area)an working area known to be free of contaminants (main tissue culture facility) ideally, tissue culture is performed in a facility specially designed for and exclusively devoted to this kind of work.- all new material should be handled as ‘quarantine material’ until it has been shown to be free of contaminants.- all work surfaces should be cleaned between activities- the potential danger of the cell material used may require additional considerations (such as the type of work surfaces and flooring, the type of microbiological safety cabinet, provision of air pressure negative to corridors)see guidelines for handling of different kinds of cell material8
اسلاید 9: EquipmentHorizontal Laminar Flow CabinetsThese provide the most sterile environment for the cells, but offer no protection to the operatorFiltered air enters at the back of the cabinet and is directed to the front, directly at the operatorThe most sterile part of the cabinet is at the back9
نقد و بررسی ها
هیچ نظری برای این پاورپوینت نوشته نشده است.