cell types.ppt

cell types.ppt

• 1.
  Lecture 47. Animalcell culture technique and types of culture s
• 2.
  Types of culturetechniques • 1. Organ culture: Blood allow to clots in a cavity and cells are grown on them. • 2. Explant culture. • 3. Disaggregation of tissue and get individual cells
• 3.
  Cell cultures aredescribed based on two parameters • Origin of the cells – Primary cell culture (from animal or plant tissue) – Extended culture (multipassage culture – cell strain) – Established (transformed) cell lines • Manner of growth – Suspension – Adherent
• 4.
  Primary cell cultures •Cells derived directly from a tissue • Limited growth potential • Limited life span – Usually cell dies after 50 divisions • Cells are used within days from culture • May give rise to a cell strain or be immortalized • Strain - a lineage of cells originating from one primary culture
• 5.
  Transformed cells, celllines • Derived from tumors • Arise spontaneously in culture • Primary cells transformed with viruses, radiation etc • Grow indefinitely in culture • Usually unstable complement of chromosomes • Cell line - cells derived from a single parental transformed cell
• 6.
  Transformed cell lines •These types of cells do not age in culture • They are ‘immortal’ • They often lose contact inhibition • They often lose many normal characteristics • They are not dependent on growth factors • They may express ‘large T-antigen’ a p53 inhibitor
• 7.
  Transformation Characteristics – Infinite lifespan – High growth potential – Low growth factor dependence – Suspension growth – Aneuploid Methods – Mutagens – Viruses – Oncogenes – Spontaneous tumors Not all transformed cell lines can form tumors, but all tumors contain transformed cells
• 8.
  Evolution of acell line
• 9.
  Advantages and Disadvantagesof finite and continuous cell lines
• 10.
  Chromosome numbers offinite and continuous cell lines
• 11.
  Classification of cellcultures based on manner of growth • Suspension cells – Grow floating in culture medium – Cells survive and proliferate without the attachment to culture container – Cells cultured from blood, spleen and bone marrow • Adherent cells – Grow as a monolayer attached to the surface of culture container – Cells derived from ectodermal and endodermal embryonic layers – Fibroblasts, epithelial cells, neurons – Most of the time require the specialized surface
• 12.
  Name Species andtissue of origin Morphology MRC-5 Human lung Fibroblast HeLa Human cervix Epithelial VERO African Green Monkey Kidney Epithelial NIH 3T3 Mouse embryo Fibroblast L929 Mouse connective tissue Fibroblast CHO Chinese Hamster Ovary Fibroblast BHK-21 Syrian Hamster Kidney Fibroblast HEK 293 Human Kidney Epithelial HEPG2 Human Liver Epithelial BAE-1 Bovine aorta Endothelial Adherent cell lines
• 13.
  Name Species and tissueof origin Morphology NSO Mouse myeloma Lymphoblastoid- like U937 Human Hystiocytic Lymphoma Lymphoblastoid Namalwa Human Lymphoma Lymphoblastoid HL60 Human Leukaemia Lymphoblastoid- like WEHI 231 Mouse B-cell Lymphoma Lymphoblastoid YAC 1 Mouse Lymphoma Lymphoblastoid U 266B1 Human Myeloma Lymphoblastoid SH-SY5Y Human neuroblastoma Neuroblast Suspension cell lines
• 14.
  BAE Endothelial HeLaepithelial MRC 5 fibroblast SHSY5 neuronal
• 15.
  Epithelial cells: Sphericalor oval, adherent, confluent monolayer is farmed After sub culturing. Strong cell adhesion and needs extracellular matrix Like dextrin, collagen, fibrin to serve as support for cell to attach and multiply. Mouse and rabbit epethilial cell lines are available from cell culture centres. Endothelial cells: Oval shaped with elongated connective fibres appear like spines Embedded in fibronectin network. Farm partial confluent monolayer after subculturing. Fibroblast cells: Rich in actin and mayosin febrile proteins. Cells appear slender and spiny with large vacuolar space between the fibrin network. Adherent cell type. Tissue specific neuronal, hepatic, pancreatoc acinar cell lines also available. Cells from spleen (lymphocytes), blood (RBC, WBC,Platelets) have their own morphology specific to their metabolic requirements.
• 16.
  • In someinstances cell cultures may grow as semi-adherent cells e.g. B95-8 where there appears to be a mixed population of attached and suspension cells. • For these cell lines it is essential that both cell types are subcultured to maintain the heterogeneous nature of the culture.