Triple gas incubator settings

[Cathy]

What are the optimal oxygen and carbon dioxide settings for triple gas
incubators used for amniotic fluid cultures?

[andrew]

Cathy the optimal oxygen is 5.0% & CO2 is 5.0%. The nitrogen will
drive the oxygen % from 20% to 5%.
U will still need to check with your vendor of the trigas incubator to
confirm these values.

[Cathy]

the vendor says this is an "operator" setting and will not
comment...thank you Andrew for your response...we thought that was
what it should be set at but just wanted to double check....

> > incubators used for amniotic fluid cultures?- Hide quoted text -
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[vantuinen]

Hee is the original citation:
Prenat Diagn. 1984 May-Jun;4(3):171-9.
The effect of oxygen tension on colony formation and cell
proliferation of amniotic fluid cells in vitro.
Held KR, Sönnichsen S.
Abstract
The effects of reduced oxygen concentrations in the gas phase over the
culture medium on colony formation and cell proliferation were
investigated in high and low cell density primary and secondary
cultures of amniotic fluid cells. Using two standard culture methods
(25 cm2 plastic flasks and Leighton type tubes) a significantly
reduced culture time was observed at high cell density for mass
cultures by incubation within a low oxygen tension gas phase (2.5 per
cent to 7.5 per cent O2) instead of conventional air (18 per cent O2).
At low cell density colony formation was significantly enhanced in
cultures grown at reduced oxygen tension. Using gas permeable
membranes as support, lowering the oxygen tension from 7.5 per cent to
2.5 per cent yielded an increase in plating efficiency of cells from
approximately 5 per cent to 25 per cent, whereas plating efficiency
was less than 2 per cent for cells grown at ambient 18 per cent O2. It
is suggested that low oxygen tension in the gas phase is an effective
means of enhancing clonal growth in amniotic fluid cell cultures,
thereby reducing both culture time and risk of culture failure.
Pete vanTuinen