The surgical reconstruction of functional neovagina is challenging and susceptible to complications. Therefore, developing tissue engineering based treatment methods for vaginal defects is important. Our aim was to develop and test a novel supercritical carbon dioxide foamed poly-l-lactide-co-ε-caprolactone (scPLCL) scaffold for vaginal reconstruction. The scaffolds were manufactured and characterized for porosity (65±4%), pore size (350±150µm) and elastic modulus (2.8±0.4MPa). Vaginal epithelial (EC) and stromal cells (SC) were isolated, expanded, and characterized with flow cytometry. Finally, cells were cultured with scPLCL scaffolds in separate and/or co-cultures. Their attachment, viability, proliferation and phenotype were analyzed. Both cell types strongly expressed cell surface markers CD44, CD73, and CD166. Strong expression of CD326 was detected with ECs and CD90 and CD105 with SCs. Both ECs and SCs attached and maintained viability on scPLCL. Further, scPLCL supported the proliferation of especially ECs, which also maintained epithelial phenotype (cytokeratin expression) during 14d assessment period. Interestingly, ECs expressed uroplakin (UP) Ia, UPIb, and UPIII markers, further, UPIa and UPIII expression was significantly higher on ECs cultured on scPLCL than on cell culture plastic. In conclusion, the scPLCL is potential scaffold for vaginal tissue engineering and the results of this study further illustrates the excellent biocompatibility of PLCL.