Increasing evidence suggests that culturing
cancer cells in three dimensions more accurately recapitulates the complexity of
tumor biology. Many of these models utilize reconstituted basement membrane derived from animals which contain a variable amount of
growth factors and
cytokines that can influence the growth of these cell culture models. Here, we describe in detail the preparation and use of
PuraMatrix, a commercially available self assembling
peptide gel that is devoid of animal-derived material and pathogens to encapsulate and propagate the
ovarian cancer cell line, OVCAR-5. We begin by describing how to prepare the
PuraMatrix prior to use. Next, we demonstrate how to properly mix the
PuraMatrix and cell
suspension to encapsulate the cells in the
hydrogel. Upon the addition of cell
culture media or injection into a physiological environment, the
peptide component of
PuraMatrix rapidly self assembles into a 3D
hydrogel that exhibits a nanometer scale fibrous structure with an average pore size of 5-200 nm(1). In addition, we demonstrate how to propagate cultures grown in encapsulated
PuraMatrix. When encapsulated in
PuraMatrix, OVCAR-5 cells assemble into three dimensional acinar structures that more closely resemble the morphology of micrometastatic nodules observed in the clinic than monolayer in vitro models. Using confocal microscopy we illustrate the appearance of representative OVCAR-5 cells encapsulated in
PuraMatrix on day 1, 3, 5, and 7 post plating. The use of
PuraMatrix to culture
cancer cells should improve our understanding of the disease and allow us to assess treatment response in more clinically predictive model systems.