Incorporation of labeled glucosamine into glycoproteins by organ cultures of hamster trachea: Adverse effects of hepes buffer

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Abstract
Hamster tracheas wer cultured in serum-free CMRL 1066 medium buffered with either NaHCO3 alone or HEPES (N-2-hydroxyethylpiperazine-N′-2-ethanesulphonic acid) plus NaHCO3 in an atmosphere of 95% O2 and 5% CO2. After 2 days in culture, tracheas maintained in HEPES plus NaHCO3-buffered medium showed an altered surface morphology. Histological examination after 6 days of culture with HEPES plus NaHCO3 showed the presence of only scattered clumps of cilia. Incorporation of [14C]-glucosamine into intracellular glycoproteins was reduced by 75% and into secreted glycoproteins by 54% in cultures buffered with HEPES plus NaHCO3, compared to NaHCO3-buffered cultures. Incorporation of [3H]fucose into intracellular glycoproteins was also reduced, although no effect was observed on secreted glycoproteins.