Thetraditional two dimensional (2D) cell culture apparatus, such as a Petri dish,is a highly synthetic environment for cell growth. Cells are three dimensional (3D) structuresand significantly alter their geometry when grown on a flat surface. This affects greatly on the ability of thecell to grow and function like its native counterparts and thus influences thedata produced from 2D culture experiments. Accordingly, there is increasingdemand to generate culture models that grow cells resembling tissues. This has led to the fabrication of 3Dscaffolds which provide the complex architecture and spatial arrangement whichallows the cells to interact with the neighboring cells in a similar way asthey would in a real tissue. In this study TERA2.cl.SP12 human embryonal carcinoma (EC) stem cells were cultured onthe 3D scaffolds and tested for comparison between the 2D and 3D cell growth,measurement of optimal seeding density and determination of cell viability andproliferation. The results suggest thatcells grown on 3D scaffolds exhibit structures and growth patterns thatresemble certain aspects of cell differentiation observed in EC stem cellderived teratomas. Different numbers ofcells were loaded onto the scaffolds and the optimal seeding density wasdetermined at1 million cells/ml. For thecell viability determination, a MTS assay was performed on different cellnumbers loaded on scaffolds and processed on different days. When 100,000 cellswere seeded into the 2D environment they initially outgrew the ones loaded onthe scaffold while the 1 million cell density loaded onto the 3D scaffoldsexhibited an exponential growth pattern backing up the observations of optimalseeding density. In conclusion, thesedata demonstrate that 3D scaffolds support the growth, expansion anddifferentiation of human pluripotent stem cells. Future experiments will test the growth ofhuman embryonic stem cells on these materials.