Replica-moulded polydimethylsiloxane culture vessel lids attenuate osmotic drift in long-term cell cultures
An imbalance in medium osmolarity is a determinant that affects cell culture longevity. Even in humidified incubators, evaporation of water leads to a gradual increase in osmolarity over time. We present a simple replica-moulding strategy for producing self-sealing lids adaptable to standard, small-...
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| Published in: | Journal of biosciences 2009-03, Vol.34 (1), p.59-69 |
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| Main Authors: | , , , |
| Format: | Article |
| Language: | eng |
| Subjects: | |
| Online Access: | Get full text |
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| Summary: | An imbalance in medium osmolarity is a determinant that affects cell culture longevity. Even in humidified incubators, evaporation of water leads to a gradual increase in osmolarity over time. We present a simple replica-moulding strategy for producing self-sealing lids adaptable to standard, small-size cell-culture vessels. They are made of polydimethylsiloxane (PDMS), a flexible, transparent and biocompatible material, which is gas-permeable but largely impermeable to water. Keeping cell cultures in a humidified 5% CO
2
incubator at 37°C, medium osmolarity increased by +6.86 mosmol/kg/day in standard 35 mm Petri dishes, while PDMS lids attenuated its rise by a factor of four to changes of +1.72 mosmol/kg/day. Depending on the lid membrane thickness, pH drifts at ambient CO
2
levels were attenuated by a factor of 4 to 9. Comparative evaporation studies at temperatures below 60°C yielded a 10-fold reduced water vapour flux of 1.75 g/day/dm
2
through PDMS lids as compared with 18.69 g/day/dm
2
with conventional Petri dishes. Using such PDMS lids, about 2/3 of the cell cultures grew longer than 30 days
in vitro
. Among these, the average survival time was 69 days with the longest survival being 284 days under otherwise conventional cell culture conditions. |
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| ISSN: | 0250-5991 0973-7138 |