Cell Culture Methods

  1. Advantages of Peripheral blood
    • Relatively fast results (3-4 days)
    • Easy to obtain
    • Can be repeated
    • can also be obtained from fetus PUBS
  2. Types of short term cell culture
    Peripheral blood and Bone marrow
  3. Long term cell cultures
    • Amniotic fluid
    • Skin biopsies and other tissues
    • Chorionic villus sampling (CVS)
  4. Peripheral blood must be collected with_____ to prevent clotting
    Sodium heparin
  5. The amount of peripharl blood collected for adults and infants
    • adults: 5-10cc
    • infants: 1/2-2cc
  6. T-lymphocytes in peripheral blood are stimulated with:
    phytohemaglutinin PHA
  7. In Peripheral blood ______ is used to detect Fragile X
    Folate (free medium or folate antagonists)
  8. In peripheral blood _______ is used to botain long chromosomes
    Methotrexate or ethidium bromide
  9. In perpheral blood BrdU (5-bromodeoxyuridine) is used for
    • DNA replication studies
    • sister chromatid exchange studies
  10. Advantages of Bone marrow
    • fast- no culture required
    • can diagnos leukemias
    • can be used for quick diagnosis in newborns
  11. ____ can be used in the diagonosis of Leukemia
    bone marrow cultures
  12. Bone marrow must be collected with ____
  13. ___ to ____ cc are required for Bone marrow cultures
    1/2 to 2
  14. Bone marrow must be processed______
  15. What is the primar use for amniotic fluid cell cultures
    prenatal diagnosis
  16. Amniotic fluid is obtained at:
    16-18 weeks gestation
  17. _____ml of fluid is required for amniotic fluid
  18. The culture time required for amniotic fluid is
    6 days to several weeks
  19. Fibroblasts (in amniotic fluid) are cultured
    • in situ converslip method
    • or
    • Flask method
  20. Alpha-fetoprotein analysis is a special treatment of
    Amniotic fluid
  21. Amniotic fluid cells can also be used for:
    biochemical or DNA testing
  22. For skin biopsies a ____ mm2 piece is obtained sterily
  23. Skin biopsies are transported in
    tissue culture medium
  24. The two methods of skin biopsy are:
    • Explant method
    • Collagenase method
  25. When taking skin biopsies are taken from autopsy specimen:
    • Skin pericaridum or other available tissue
    • must be obtained quickly and sterily
    • must avoid exposure to fixatives
    • transport in tissue culture medicum
  26. Most cell culture methods involve the culture of a cell called ______ which can be proliferated for up to 50 or 60 cell doublings
  27. Chorionic villus sampling is used to:
    to obtain a sufficient quant of living tissue for direct DNA extraction without the use of a cell culture

    early prenatal detection of chromosome abnormalities
  28. Methods also used for early prenatal detecton of chromosome abnormalities:
    • direct chrom analysis of trophoblasts
    • through long term culture of mesenchymal cores
  29. CVS is typically obtained at _____gestation
    9-11 weeks
  30. Risks of CVS include:
    fetal limb defects due to disruption of the blood suppy
  31. _______ may also be a complicating factor in prenatal chromosome analysis
    Tissue Specific Mosaicism
  32. Phytohemagglutinin (PHA) is a protien isolated from the common ______
    Red kidney bean
  33. PHA caused agglutination of _______ by interacting with cell surfaces
    T lymphocytes
  34. ________ is a natural alkaloid from a flowering crocus plant, not typically replaced with _________

  35. Sodium heparin
    is a natural anticoagulant that prevents the clotting of blood
Card Set
Cell Culture Methods
Cytogenetics midterm