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Advantages of Peripheral blood
Relatively fast results (3-4 days)
Easy to obtain
Can be repeated
can also be obtained from fetus PUBS
Types of short term cell culture
Peripheral blood and Bone marrow
Long term cell cultures
Amniotic fluid
Skin biopsies and other tissues
Chorionic villus sampling (CVS)
Peripheral blood must be collected with_____ to prevent clotting
Sodium heparin
The amount of peripharl blood collected for adults and infants
adults
: 5-10cc
infants
: 1/2-2cc
T-lymphocytes in peripheral blood are stimulated with:
phytohemaglutinin PHA
In Peripheral blood ______ is used to detect Fragile X
Folate (free medium or folate antagonists)
In peripheral blood _______ is used to botain long chromosomes
Methotrexate or ethidium bromide
In perpheral blood BrdU (5-bromodeoxyuridine) is used for
DNA replication studies
sister chromatid exchange studies
Advantages of Bone marrow
fast- no culture required
can diagnos leukemias
can be used for quick diagnosis in newborns
____ can be used in the diagonosis of Leukemia
bone marrow cultures
Bone marrow must be collected with ____
heparin
___ to ____ cc are required for Bone marrow cultures
1/2 to 2
Bone marrow must be processed______
immediately
What is the primar use for amniotic fluid cell cultures
prenatal diagnosis
Amniotic fluid is obtained at:
16-18 weeks gestation
_____ml of fluid is required for amniotic fluid
20-30ml
The culture time required for amniotic fluid is
6 days to several weeks
Fibroblasts (in amniotic fluid) are cultured
in situ converslip method
or
Flask method
Alpha-fetoprotein analysis is a special treatment of
Amniotic fluid
Amniotic fluid cells can also be used for:
biochemical or DNA testing
For skin biopsies a ____ mm2 piece is obtained sterily
5-10
Skin biopsies are transported in
tissue culture medium
The two methods of skin biopsy are:
Explant method
Collagenase method
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
Most cell culture methods involve the culture of a cell called ______ which can be proliferated for up to 50 or 60 cell doublings
Fibroblast
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
Methods also used for early prenatal detecton of chromosome abnormalities:
direct chrom analysis of
trophoblasts
through long term culture of
mesenchymal cores
CVS is typically obtained at _____gestation
9-11 weeks
Risks of CVS include:
fetal limb defects due to disruption of the blood suppy
_______ may also be a complicating factor in prenatal chromosome analysis
Tissue Specific Mosaicism
Phytohemagglutinin (PHA) is a protien isolated from the common ______
Red kidney bean
PHA caused agglutination of _______ by interacting with cell surfaces
T lymphocytes
________ is a natural alkaloid from a flowering crocus plant
, not typically replaced with _________
Colchicine
Cocemid
Sodium heparin
is a natural anticoagulant that prevents the clotting of blood
Author
minna528
ID
21865
Card Set
Cell Culture Methods
Description
Cytogenetics midterm
Updated
2010-06-03T13:21:46Z
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