An hypothesis on the role of cellular colloid osmotic pressure in determining behavior of cells in vitro including anchorage dependency and maintenance of the differentiated state.

The osmotic problems involved when cells are isolated from tissues are analyzed. Evidence is considered which indicates that in vivo the Na pump is operating at maximal or near maximal rates and that this depends on low leak rates for salts and water due to various aspects of the tissues structure. Dispersion of the tissue results in breakdown of these barriers on free diffusion and the isolated cell is subjected to an enormous increase in passive influx due to colloid osmotic pressure without being able to increase its pumping rate to the extent needed to maintain volume control. It is proposed that the primary problem the cell faces in vitro is to compensate for the effective increase in its colloid pressure, e.g. the colloid osmotic pressure excess, emerging with the breakdown of the tissue structure. The finding that most normal cells have to adhere to a surface in order to grow or "anchorage dependency" is analyzed in terms of the way adhesion and spreading result in changes in ion and water movements into cells enabling them to achieve fluid balance in the face of the colloid pressure excess. It is also proposed that the differentiated state is more dependent on colloid osmotic balance than proliferation. The failure of conditions used in tissue culture to compensate adequately for the colloid pressure excess results in limiting the amount of protein which can be synthesized, dissipation of cellular energy, and changes in orientation of cellular components which contribute directly to the loss of differentiation which occurs during growth in vitro.