Role of cell density/cell‐cell contact, and growth state in expression of differentiated properties by the LLC‐PK1 Cell

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Abstract
Populations of the renal epithelial cell line, LLC‐PK,1, acquire many properties characteristic of the proximal tubular cell at confluence. At confluence cells both enter a nonproliferative state and develop extensive cell‐cell contacts. To determine if one or both factors is responsible for acquisition of the differentiated phenotype, growth arrest was initiated in populations of varying densities by two procedures (serum deprivation and thymidine block) and expression of several differentiated properties (Na‐hexose symport activity, gamma‐glutamyl transpeptidase activity, alkaline phosphatase activity, and villin protein) was examined. Induction of growth arrest resulted in expression of all differentiated properties even in subconfluent populations. The level of expression in a population was proportional to cell density at the initiation of growth arrest; higher density was associated with increased expression. Evidence indicated the existence of some minimal density below which cells could not express detectable levels of differentiated properties in response to induction of growth arrest. The procedure used to initiate growth arrest did not affect this behavior, indicating that initiation of cell growth arrest rather than hormone deprivation was the inducing factor. These results indicate that both cell growth state and cell density independently modulate expression of differentiated properties by the LLC‐PK1 cell. These results are incorporated into a model in which cells in the absence of “appropriate” cell‐cell contact arrest at a differentiation‐incompetent cell cycle point. In the presence of appropriate cell‐cell contact (as yet undefined) cells arrest at a distinct differentiation‐competent cell cycle point and initiate expression of the differentiated phenotype.

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