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Ascites fluid collection

Mar 30th 2015 at 11:23 PM

Guideline: Depending on the condition
of the mouse, a maximum of two taps of
the ascitic fluid are permitted, with the
second tap being a terminal procedure.
Training and experience in tapping or
draining ascitic fluid is essential.
It should be noted that fluid removal carries
the risk of hemorrhage, oedema and death.
For more aggressive cell lines, known to
cause significant morbidity, the number of
taps should be limited to one terminal
procedure under general anesthesia.
Anesthesia, as recommended by current
CCAC guidelines may be used for the first
(non-terminal) tap. However, anesthesia
leads to a decrease in blood pressure and
respiratory suppression, which may be
detrimental to the already compromised
mouse. Therefore, it is recommended that
mice are placed in an oxygen filled
induction chamber for 5-10 minutes,
followed by mild gaseous anesthesia
(isoflurane) to give good immobilization
of the animal for the procedure with rapid
recovery. To minimize any bacterial contamination,
the site of paracentesis should be
aseptically prepared for the first tap.
It is recommended that a large gauge
needle (21-22 gauge) be used for paracentesis.
A large gauge needle permits rapid
collection of the viscous ascitic fluid, reducing
the period of restraint/anesthesia
required; however, too large a needle can
cause tissue damage. A maximum of 4-5ml
of ascitic fluid may be collected at the first
(survival) tap. The abdomen of the mouse
should be palpated to determine whether an
intra-abdominal solid tumor is present,
the presence of a tumor being grounds
for humane euthanasia of the animal.
Administration of replacement fluids (1-2ml
subcutaneous) should be considered when
large volumes of ascitic fluid are harvested
and the animals are not terminated
(Jackson & Fox, 1995).
The animals should be closely monitored for
the first 60 minutes and regularly for several
hours following the first tap. Any signs of
distress should result in euthanasia of the
animal, according to current CCAC guidelines.
ACCs must ensure that the personnel
responsible for carrying out these procedures
(listed on the animal use protocol)
have obtained the necessary training
prior to conducting the procedure. CCAC
guidelines on: institutional animal user
training (1999) mandates the training of any
personnel prior to the performance of
animal-based procedures.



Related Tags : antibody production , production of monoclonal antibodies

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