We have developed a very successful process of running Sp2 cells in 250mL shake flask, 2L bioreactor, 5L bioreactor, in 10L bioreactor for the last two years. The cultures lasted 12 - 14 days. Recently, i cannot repeat the process. The reasons, i don't know why the NH4 in the culture accumulated too high (~ 8 mM compared with 5mM previously). Cells kept growing at 32C (temperature shift from 37 to 32 on D4). Cell had a big drop of viability on D6, D7 (20% vs 10% previously). Then it could not last till D12 as previously.
All the parameters were kept the same. Cell source the same. Media, the same. What is, are the possible reason of failure of repeat the process?
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