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Role of Angiogenesis in Diseases

Objectives
Activities
Questions

Objectives [top]

  1. Discuss hematopoietic cells and the bone marrow microenvironment, and:
    1. assays to measure hematopoietic progenitors;
    2. assays to measure endothelial cell (EC) functions (morphogenesis, proliferation).
  2. Discuss involvement of endothelial cells in cancer development – tumor angiogenesis.
  3. Aseptically harvest, centrifuge, resuspend cells in Matrigel or methylcellulose.
  4. Discuss the use of specific growth factors for differentiation of a specific lineage.
  5. Discuss formation of hematopoietic colonies, endothelial cell morphogenesis (in vitro capillary-like structures), formation of cellular networks and 3D structures.
  6. Discuss use of Matrigel assay for screen for anti-angiogenic drugs.

Activities [top]

  1. Observe EC monolayer under microscope and discuss how EC were isolated.
  2. Learn sterile techniques used for tissue culture preparation.
  3. Aseptically harvest ECs from plates provided. Cells will be placed in tubes and centrifuged.
  4. Discard media, resuspend cell pellet, and count cells. Resuspend cells in Matrigel and plate them into a 48 well tissue plate.
  5. Observe capillary-like structures formed in Matrigel using the microscope, and take pictures; observe the effect of specific drug on capillary-like structure formation.

Questions [top]

  1. Why do EC display a different behavior in Matrigel than when cultured in plastic?
  2. Which properties distinguish EC cells from the other adherent cell lineages in Matrigel?
  3. What growth factors are necessary to support EC functions?
  4. Why use drugs that inhibit specific signaling molecules to inhibit EC and tumor angiogenesis?