Chinese hamster lymphocyte cultures. Relationship between lymphocyte proliferation,cell concentration,culture time and culture area

Different numbers of Chinese hamster lymphocytes were cultured in microtiter plates with flat-, round- and V-bottomed wells for different culture times. The smallest number of cells could be stimulated in plates with V-bottomed wells. At least 25,000–50,000 cells and a longer culture time than for round- or V-bottomed plates were required for maximal stimulation in flat-bottomed plates.For a given well conformation the optimal day of culture is earlier with higher and later with lower cell concentrations.The optimal culture time for a given number of cells is shortest in V- and longest in flat-bottomed plates.The amount of PHA producing the highest thymidine incorporation for a given number of cells depends upon the well conformation. It decreases with lower cell concentration and increases with longer culture time.