The Sansevieria plant is widely appreciated for its upright, fleshy, and attractive leaves, as well as its high adaptability to home conditions. Due to the presence of preclinal chimera tissue in this species and the challenges associated with propagating it via leaf cuttings, finding an effective method to enhance the production of various variegated Sansevieria cultivars is essential. In this study, we examined the longitudinal thin cell layer technique for producing mutated Sansevieria varieties. Our results indicated that plantlets regenerated directly or indirectly from the epidermal layers of the chimeric Sansevieria were, contrary to expectations that they would resemble the mother plant, yellowish mutants, with none resembling the mother plant. In indirect regeneration, shoots were formed from the callus after callus induction. Samples grown in 2,4-D medium were subsequently subcultured in the same medium and showed regeneration without the need for a cytokinin after 80 days. After 80 days of culturing, some samples rooted without requiring IBA. The highest average number of leaves and plantlets in indirect regeneration occurred in the MS medium containing 0.6 mg/L of 2,4-D, where approximately 2.5 plantlets were produced after 80 days from the callus. In contrast, the 2,4-D concentration of 0.1 mg/L produced about 1.5 plantlets. The MS medium containing 1.2 mg/L of BA led to the highest direct shoot regeneration (3.3) using the thin cell layer culture technique. Only 65% of the regenerated samples were able to acclimatize after rooting four months later. Our investigation indicates that producing a preclinal chimera plant resembling the mother plant using the longitudinal thin cell layer culture technique from the outer epidermal layers is not feasible; nearly all produced plants turned out to be yellowish mutants, significantly different from the typical green plants. This finding is noteworthy as no previous studies have investigated the production of variegated ornamental cultivars through thin cell layer culture methods. The results of this research could provide valuable insights for the future production of variegated plant varieties via in vitro culture.