Animal Cell and Tissue Culture Lecture 1 - 2024-2025

Animal cell and tissue culture Lecture 1- Dr Seham Aly 2024-2025  ‫رؤية ورسالة كلية العلوم‬ ‫رؤية الكلية‬ ‫ ‬ ‫التمٌز فً تعلٌم العلوم األسـاسٌة والبحث العلمً للمساهمة فى التنمٌة المستدامة‪.‬‬ ‫ ‬...

Animal cell and tissue culture Lecture 1- Dr Seham Aly 2024-2025  ‫رؤية ورسالة كلية العلوم‬ ‫رؤية الكلية‬ ‫ ‬ ‫التمٌز فً تعلٌم العلوم األسـاسٌة والبحث العلمً للمساهمة فى التنمٌة المستدامة‪.‬‬ ‫ ‬ ‫رسالة الكلية‬ ‫ ‬ ‫تقدٌم تعلٌم ممٌز فً مجاالت العلوم األساسٌة و إنتاج بحوث علمٌة تطبٌقٌة للمساهمة فى‬ ‫ ‬ ‫التنمٌة المستدامة من خالل إعداد خرٌجٌن متمٌزٌن طبقا للمعاٌٌر األكادٌمٌة القومٌة‪ ،‬و‬ ‫تطوٌر مهارات و قدرات الموارد البشرٌة‪ ،‬و توفٌر خدمات مجتمعٌة وبٌئٌة تلبً طموحات‬ ‫مجتمع جنوب الوادي‪ ،‬و بناء الشراكات المجتمعٌة الفاعلة‪.‬‬ Course content BTC 403–Cell and Tissue culture (Animal) Introduction to tissue culture, General methodologies in tissue culture, Media components, preparation, and contamination, Cryopreservation, Somatic embryogenesis, Physiology of cell culture, animal Tissue culture part: Animal Cell Culture: Historical Background - Importance of and progress in Animal Cell Culture Technology - Biology of animal Cell; basic concepts (Cellular Interactions) - Culture media of animal cells: Serum and Serum Free Media - Cell metabolism and its control in culture media - Mechanism of cell proliferation and cell death in vitro - Culturing and Sub-Culturing of Animal Cells - Monitoring and control of cell culture - Cell Differentiation & Cell Movement - Cloning of Animal Cells - Cell Line Preservation- Cell Line Characterization- Mycoplasma: Detection and Control - Monoclonal Antibody Production - product purification processes. Course syllabus  Cell culture refers to: The removal of cells from an animal or plant and their subsequent growth in a favorable artificial environment closely resembles in vivo Cells and tissues. Or 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. Historical events in the development of cell culture Why are cell culture used for/ applications of cell and tissue culture? 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. 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. Types of cell cultures  Resected Tissue Cell or tissue culture in vitro Primary culture Organ culture Primary cell culture Slice tissue culture Tissue culture Sub-culture Secondary culture Sub-culture Continuous culture (cell line) Transformed cell lines normal cell line Neoplastic cell lines Stem cells Hybrid cell line How are cell cultures obtained? There are three methods commonly used to initiate a culture from animals. 1- Organ culture Whole organs from embryos or partial adult organs are used to initiate organ culture in vitro. These cells in the organ culture maintain their differentiated character, their functional activity, and also retain their in vivo architecture. They do not grow rapidly, and cell proliferation is limited to the periphery of the explant. As these cultures cannot be propagated for long periods, a fresh explanation is required for every experiment that leads to interexperimental variation in terms of reproducibility and homogeneity. 2-Primary explant culture Fragments exercised from animal tissue may be maintained in a number of different ways. The tissue adheres to the surface aided by an extracellular matrix (ECM) constituent, such as collagen or a plasma clot, and it can even happen spontaneously. This gives rise to cells migrating from the periphery of the explant. 3-Cell culture This is the most commonly used method of tissue culture and is generated by collecting the cells growing out of explants or dispersed cell suspensions (floating free in culture medium). Cells obtained either by enzymatic treatment or by mechanical means are cultured as adherent monolayers on solid substrate. Types of cell cultures Cell culture is of three types: (1) precursor cell culture, which is undifferentiated cells committed to differentiate; (2) differentiated cell culture, which is completely differentiated cells that have lost the capacity to further differentiate; and (3) stem cell culture, which is undifferentiated cells that go on to develop into any type of cell. Cells with a defined cell type and characteristics are selected from a culture by cloning or by other methods; this cell line becomes a cell strain. Monolayer cultures The monolayer culture is an anchorage-dependent culture of usually one cell in thickness with a continuous layer of cells at the bottom of the culture vessel. Suspension cultures Some of the cells are nonadhesive and can be mechanically kept in suspension, unlike most cells that grow as monolayers (e.g., cells of leukemia). This offers numerous advantages in the propagation of cells. General concepts Primary culture refers to the stage of the culture after the cells are isolated from the tissue and proliferated under the appropriate conditions until they occupy all of the available substrate (i.e., reach confluence). At this stage, the cells have to be subcultured (i.e., passaged) by transferring them to a new vessel with fresh growth medium to provide more room for continued growth. Cell Line After the first subculture, the primary culture becomes known as a cell line or subclone. Cell lines derived from primary cultures have a limited life span (i.e., they are finite; see below), and as they are passaged, cells with the highest growth capacity predominate, resulting in a degree of genotypic and phenotypic uniformity in the population. Cell Strain If a subpopulation of a cell line is positively selected from the culture by cloning or some other method, this cell line becomes a cell strain. A cell strain often acquires additional genetic changes subsequent to the initiation of the parent line. Finite vs Continuous Cell Line Normal cells usually divide only a limited number of times before losing their ability to proliferate, which is a genetically determined event known as senescence; these cell lines are known as finite. However, some cell lines become immortal through a process called transformation, which can occur spontaneously or can be chemically or virally induced. When a finite cell line undergoes transformation and acquires the ability to divide indefinitely, it becomes a continuous cell line. Why sub culturing? ** Subculturing: the removal of the medium and transfer of cells from a previous culture into fresh growth medium. ** Confluency: Is the term commonly used as a measure of the number of the cells in a cell culture dish or flask, and refers to the coverage of the dish or the flask by the cells. Once the available substrate surface is covered by cells (a confluent culture) growth slows & ceases. Cells to be kept in healthy & in growing state have to be sub-cultured or passaged. It’s the passage of cells when they reach to 80-90% confluency in flask/dishes/plates The passage number is the number of sub-cultures the cells have gone through. Passage number should be recorded and not get too high. Cell lines with passage numbers of greater than 30 are more likely to acquire genetic abnormalities compared to lower passage cells. Next lecture THANK YOU!