Liquid nitrogen (LN) storage is recommended for conserving plants with seeds for which standard seed banking is not feasible, but there are few empirical reports confirming its long-term effectiveness. In this study we evaluated in vitro viability and ex vitro growth of embryo axes of a species that is short-lived in seed bank conditions (Juglans nigra) and two recalcitrant seeded species (Aesculus hippocastanum and A. glabra) that were stored 11-23 years in LN. Viability of J. nigra axes did not decrease significantly after 23 years and produced normal-appearing plantlets. Similarly, viability of A. hippocastanum axes did not decrease after 23 years. However, A. glabra axes showed a significant decline in viability after 23 years, from 80% when freshly harvested to 33%. These results demonstrate that LN storage can preserve embryo axes of J. nigra and Aesculus sp. for over two decades, providing a workable conservation tool for these and similar species.