Asplenium scolopendrium var. americanum is a federally-listed fern species that is found in several northern populations in the U.S. and Canada, as well as in three isolated locations in the southern U.S. In vitro methods were applied to the germination, propagation, and cryopreservation of this species. Germination of spores was obtained both in vitro and on soil, but the production of sporophytes from gametophytes did not begin until over two years after the spores were sown. In vitro cultures of both sporophytes and gametophytes were established, but sporophyte production from gametophytes has thus far only been observed on soil. Both in vitro-grown gametophytes and sporophytes survived cryopreservation using the encapsulation-dehydration procedure. Preculture on ABA-containing medium increased survival of sporophytes after cryopreservation, but good survival of gametophytes was observed with or without ABA. The use of Green Globular Body-like sporophyte tissue demonstrates the use of this tissue for fern cryopreservation. The success of these protocols demonstrates that in vitro methods can be applied to the propagation and preservation of A. scolopendrium var. americanum, and possibly other fern species with similar growth forms. Propagated plants could assist in restoration of the species and cryopreservation could be used to preserve germplasm of such propagated lines into the future.