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Primary and Secondary Cell Line

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Its all about cell lines and cell cultures.

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Primary and Secondary Cell Line

  1. 1. CELL • It is the smallest unit of life. • It is the basic structural, functional, and biological unit of all known living organisms. • It is consisted of cytoplasm enclosed within a membrane, which contains many biomolecules such as proteins and nucleic acids. • First discovered by Robert Hooke in 1665. • It has a dimensions between 1- 100 micrometres.
  2. 2. CELL LINE • Cells of a multicellular organism. • The cells are grown for prolonged periods in vitro. • First discovered by Dr. George Otto Gey in 1951. • First discovered cell line, HeLa cell line. • Some cell lines are known as immortalized cell line. • Cell lines are a very important tool for research.
  3. 3. CLASSIFICATION Cell Line By Appearance Finite Cell Line Continuous Cell Line By Culture Primary Cell Line Secondary Cell Line
  4. 4. BY APPEARANCE Finite Cell Lines: • Cell Lines where cell division occurs for finite time period. • Usually represents the losing ability of proliferation of the cultured cell. • It is determined by genetic event called “senescence”. Continuous Cell Lines: • Cell Lines where cell division occurs indefinitely also known as “immortalized cell line”. • Cultured cells are usually spontaneously, chemically or virally induced. • Usually neoplastic.
  5. 5. BY CULTURE Primary Cell Line: • Derived directly from animal tissue • May be normal or neoplastic. • Cultured either as tissue explants or single cells. • Cultured in vitro. • Retain differentiated phenotype. • Mainly anchorage dependent. • Exhibit contact inhibition. • Not viable for long time.
  6. 6. SECONDARY CELL LINE: • Done by sub-culturing of primary cell line. • Usually by transferring of cells from one culture vessel to another. • Viable for long time. • Anchorage independent. • Consisting of proliferating cells. • Cell line gets continuously evolved and properly established.
  7. 7. CELL CULTURE • Cells are grown under controlled conditions (in vitro). • Cells isolated from living tissues. • Carried out under controlled conditions like essential nutrients, growth factors, hormones, and gases. • Artificial substrate (adherent or monolayer culture) or culture medium (suspension culture) are required for it.
  8. 8. CLASSIFICATION Cell Culture Primary Culture Adherent Culture Suspension Culture Secondary Culture
  9. 9. IN-VITRO CELL CULTURE Animal Cell Culture: • Used provide a good model system for studying. • Used in toxicity testing. • Used to observe replication of viruses in cell cultures . Plant Cell Culture: • Used to produce clones of a plant. • To quickly produce mature plants. • Mainly based on totipotency. • Large scale production of artificial seeds.
  10. 10. COMMON EXAMPLES Cell Line Organism Origin Tissue HeLa Human Cervical Cancer 293-T Human Kidney (Embryonic) A-549 Human Lungs Carcinoma ALC Mouse Bone Marrow CHO Hamster Ovary FM 3 Human Metastatic Lymph Node Vero Monkey Kidney Caco-2 Human Colo-rectal Carcinoma MCF-7 Human Breast Tumor
  11. 11. HeLa Cells 293-T Cells A-549 Cells ALC Cells CHO Cells Vero Cells Caco-2 Cells MCF-7 Cells FM-3 Cells
  12. 12. CELL LINE MAINTENANCE • Creating master or working cell banks. • Before use, cells must be quickly thawed to prevent crystallization. • Transfer cells in tissue culture to perform cell counting. • Incubation in 370C with 5% CO2 in 10% FBS. • For sub-culturing cells are trypsinized. • Storage in 10% DMSO and 10% FBS at -200C for 1hr. and then at - 800C and then cryopreserved.

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