It has been established that although an inhibitory interaction occurs when a culture of human diploid cells become crowded together (contact inhibition of growth), multiple-layered cell sheets are obtained by using a continuous medium perfusion culture. A similar effect is obtained when the culture medium is changed at frequent intervals, and this paper reports the effects of a medium change on cell growth and metabolism.
A direct relationship was found between cell yield and the number of medium changes given to a culture. This was an unexpected result because normally when a culture is prolonged by additional feeding the cell yield shows a diminishing return. The amino acid and glucose uptakes and growth yields (the ratio of the amount of cell dry weight produced to substrate used) were determined and they also showed that a unit amount of growth occurred per medium change, and that cessation of growth was accompanied by cessation of nutrient uptake and metabolism. Medium changes had a profound affect on cellular metabolism, especially on DNA and protein synthesis. As a culture approached confluency, DNA, RNA and protein synthesis were sequentially inhibited. After a medium change there was a sequential stimulation of DNA, RNA and protein synthesis in the same order as they were inhibited. The inhibitory mechanism that is affected by cell crowding is obviously reversed by a medium change.
The results presented in this paper suggest that contact inhibition of growth primarily affects DNA synthesis and that if the cell is able to take up a sufficient supply of nutrients in a crowded culture then this inhibition can be overcome.