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Brief Exposure to Cold Culture Media Does not Affect Erythroid
Proliferation and Differentiation
Chanatip Metheetrairut, Saiphon Poldee, Kongtana Trakarnsanga*
Department of Biochemistry, Faculty of Medicine, Siriraj Hospital, Mahidol University
*Corresponding Author E-mail: kongtana.tra@mahidol.ac.th
Background: Abstract
It is common practice to keep cell culture media refrigerated and repeatedly warm them
before every medium change. However, cell culture media contain amino acids, sugars, and
vitamins as well as other proteins that may be sensitive to temperature fluctuations; and
it has been shown that temperature of fetal bovine serum storage affected some cell lines’
viability. Erythroid cell culture uses a defined cell culture media and cytokines. Although
prolonged hypothermic stress (4-24 hours, depending on cell types) has been shown to
lead to decreased cell viability, here we hypothesize that a short period of exposure to low
temperature does not affect erythroid cell proliferation, viability, or morphology.
Methods: Results: Erythroblast cell lines were cultured in an expansion medium for 2 days, following by
a differentiation medium for 7 days in an incubator at 37oC with 5% CO
2 except during
treatment to allow observation of both the cell expansion and differentiation stages. The
group receiving cold treatment was moved to a 4oC refrigerator for 1 hour daily. Cell
proliferation, viability, and morphology were observed with Trypan Blue and Leishman’s
staining.
The proliferation and cell viability rates of the two groups of cells were not significantly
different. Cell morphology was also similar between the control and the group briefly
exposed to cold cell culture media when examined after being differentiated for 7 days.
Conclusion: A short period of exposure to cell culture media at refrigerated temperature (4oC) does not
affect erythroid cell proliferation and viability as well as erythroid differentiation. Further
study may be able to demonstrate whether there are differences in the effect of cytokines or
nutrients after temperature fluctuations over time.
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