Tuxpeño is a Mexican maize landrace adapted to a wide range of environments and is agriculturally and economically relevant worldwide. To contribute to the preservation of the Tuxpeño germplasm and to develop a platform that allows performing biotechnological improvements of its traits for human benefit, this study aims to explore the potential of Tuxpeño maize to produce in vitro embryogenic callus tissue with plant regeneration capability via somatic embryogenesis. When compared to CML137, a maize line well known for its considerable plant regeneration capability, the immature Tuxpeño embryos produced embryogenic calli that efficiently proliferated and regenerated plantlets through somatic embryogenesis. Importantly, a microscopic analysis of the somatic embryogenesis process mirrored the major histological events of zygoticembryogenesis from a unicellular origin. The amount of Tuxpeño callus tissue generated on N6 medium, the number of regenerated plants from these cultures and the plant regeneration percentage were not affected by silver nitrate (AgNO₃), and these parameters were found to be comparable between the Tuxpeño and CML137 cultures. The callus yield, callus proliferation, and number of regenerated plants from the Tuxpeño embryogenic cultures were enhanced with cocktail 20 (ck20) and 2,4-dichlorophenoxyacetic acid (2,4-D) but not with 3,6-dichloro-2-methoxybenzoic acid (Dicamba) or 2-(2 methyl, 4-chlorophenoxy propionic acid (MCPP). Therefore, we established the conditions for successful plant regeneration via somatic embryogenesis from a callus culture of the maize landrace Tuxpeño. This achievement offers the opportunity to preserve the great genetic resource represented by the Tuxpeño germplasm and to improve the genotype through molecular biotechnology tools for human benefit.