Abstract: The present invention is based in part on the discovery of significant improvements to cell culture systems and methods in terms of utility and range of application. The system allows for culture of cells in one or more defined spaces within a culture chamber defined by one or more spaced opposing surfaces to provide significant advantages over previously described culture methods. The system includes a chamber for receiving a fluid culture medium; a first surface and one or more second surfaces defining one or more spaces in the chamber; and one or more cells constrained within the one or more spaces. The second surface includes at least one opening for passage of a substance into the culture space.