Stationary cultures monolayers

The cultivation of hepatocytes in a stationary suspension culture is actually ineffective. The hepatocytes lose their differentiation within hours. An improvement was the attachment culture. Thereby, the cells are cultivated either in self- or microcarrier-induced multicellular aggregates or on membranes. When standard monolayer culture was adequate to maintain the cell viability for 1 to 2 weeks the differentiation was lost after a few days. Different modifications as described beneath allowed the maintenance of differentiation for 2 to 3 weeks. 

After 3-5 days the culture density will typically reach 1-2 x 10 mL (for many hybridomas, 0.8-1.5 x 10 mH only) and can be harvested, or the culture prolonged for extra cell growth with twice-daily 50% medium changes. Suspension cells tend to have only a limited stationary phase and it is thus important to monitor growth more closely than in monolayer culture to ensure that healthy, rather than dying/dead, cells are harvested. 

As the first step in scale-up is (or should he) to produce enough cells to lay down a cell bank in liquid nitrogen Protocol 2 describes the procedure for growing cells in stationary monolayer cultures with the purpose of creating a cell bank. 

Growth of hybridoma cells in stationary monolayer cultures... 

Growth of hybridoma cells in stationary monolayer cultures 130 Growth of hybridoma cells in Nunc Cell Factories 131... 

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