Stem cell plays a significant role in tissue engineering and regenerative medicine. However, one of the major limitations in translation of stem cell technologies for clinical applications is limited cell survival and growth upon implantation. To address this limitation, authors have made an attempt to design polyacrylamide/alginate (PAAm/Algi) based tough hydrogel substrates and studied their impact on the survival and proliferation of human bone marrow-derived mesenchymal stem cells (hBMSCs). The PAAm/Algi hydrogel substrates have been prepared by initiator-induced free radical polymerization with mechanical properties quite similar to human soft tissues. To evaluate the efficacy of hydrogel substrates in support of cellular functions, hBMSCs were cultured on the PAAm/Algi hydrogel substrate (Gel system) and conventional tissue culture plate (TcP system) under defined conditions. The results of this study demonstrated that the cells cultured on the Gel and TcP systems showed 80–90% of cell viability throughout the period of study. The cells cultured on the Gel system showed 25% increase in proliferation after 7 days of culture, whereas the TcP system showed only an increase of 10%. These results confirm the cellular compatibility and enhanced cell proliferative nature of the hydrogel substrates, due the fact that the hydrogel substrates provided necessary microenvironmental cues to the cells as compared the conventional TcP system. The overall results suggest that the PAAm/Algi based hydrogels could be used as a potential substrate for hBMSCs culture and expansion. © 2017 Elsevier B.V.