Simulating Properties of In Vitro Epithelial Cell Morphogenesis

How do individual epithelial cells (ECs) organize into multicellular structures? ECs are studied in vitro to help answer that question. Characteristic growth features include stable cyst formation in embedded culture, inverted cyst formation in suspension culture, and lumen formation in overlay culture. Formation of these characteristic structures is believed to be a consequence of an intrinsic program of differentiation and de-differentiation. To help discover how such a program may function, we developed an in silico analogue in which space, events, and time are discretized. Software agents and objects represent cells and components of the environment. “Cells” act independently. The “program” governing their behavior is embedded within each in the form of axioms and an inflexible decisional process. Relationships between the axioms and recognized cell functions are specified. Interactions between “cells” and environment components during simulation give rise to a complex in silico phenotype characterized by context-dependent structures that mimic counterparts observed in four different in vitro culture conditions: a targeted set of in vitro phenotypic attributes was matched by in silico attributes. However, for a particular growth condition, the analogue failed to exhibit behaviors characteristic of functionally polarized ECs. We solved this problem by following an iterative refinement method that improved the first analogue and led to a second: it exhibited characteristic differentiation and growth properties in all simulated growth conditions. It is the first model to simultaneously provide a representation of nonpolarized and structurally polarized cell types, and a mechanism for their interconversion. The second analogue also uses an inflexible axiomatic program. When specific axioms are relaxed, growths strikingly characteristic of cancerous and precancerous lesions are observed. In one case, the simulated cause is aberrant matrix production. Analogue design facilitates gaining deeper insight into such phenomena by making it easy to replace low-resolution components with increasingly detailed and realistic components.Synopsis: To gain new insights into how normal and abnormal epithelial cell (EC) morphogenesis occurs, Grant and colleagues designed, built, and studied a series of discrete event analogues capable of mimicking epithelial growth characteristics in four different culture conditions. The analogues use independent software agents and objects to represent cells and the two environment components. “Cells” interact with local components using an axiomatic decisional process deduced from experimental in vitro observations. During simulations, “cells” form stable structures that mimic counterparts in cell cultures: a set of targeted in vitro phenotypic attributes is matched by the analogue's phenotype. However, the foundational analogue failed to exhibit a behavior characteristic of functionally polarized ECs in stable structures. Iterative refinement solved the problem: the improved analogue is the first model to simultaneously provide a representation of nonpolarized and structurally polarized cell types, and a mechanism for their interconversion. Inflexible axiom application is essential to simulate normal attributes. Selectively changing an axiom or relaxing its application caused growths strikingly characteristic of cancerous and precancerous lesions. Gaining deeper insight into such phenomena can be achieved by replacing low-resolution components with increasingly detailed and realistic components.