Nonwoven polyethylene teraphathalate (PET) fabrics vi with different porosities and knitted fabric were used as support matrixes to grow human trophoblast cells to study the spatial effects of fibrous matrix on cell adhesion, spatial organization, proliferation, and metabolic functions. In general, cells grown on 2-D surface and knitted fabric had faster metabolic rates and also showed higher proliferation activities as detected by cyclin B assay. For nonwoven PET fibers, matrix porosity had profound effects on cell morphology, spatial organization, and proliferation. Cells grown in a low-porosity fibrous matrix formed small aggregates (similar to 100 cells per aggregate), whereas cells grown in high-porosity matrix formed big aggregates (similar to 1000 cells per aggregate). This was attributed to the difference in pore volume or averaged fiber distance, which dictated a cell＇s ability to cross over and form a bridge between adjacent fibers. The high-porosity matrix had a relatively poor surface accessibility for cells to attach and spread, which are essential for cell proliferation. Dual staining with PI and BrdU showed that 60% of cells in the small aggregates found in the low-porosity matrix were proliferating, while only 18% of cells in the large aggregates found in the high-porosity matrix were proliferating. These results suggest that spatial characteristics of fibrous matrix are important to cell proliferation and function and should be considered in tissue-engineering human cells.