The germination and micropropagation of Asparagus officinalis L. remain understudied, despite its nutritional and medicinal value. This study focused on the influence of plant growth regulators (PGRs) 6-benzylaminopurine (BAP), naphthalene acetic acid (NAA), and 2,4-dichlorophenoxyacetic acid (2,4-D) on in vitro propagation efficiency. Two approaches were used: seed germination and micropropagation via in vitro nodal explants were cultivated on MS medium with varying concentrations of BAP (0.5, 1, or 1.5 mg/L), either individually or combined with 1 mg/L NAA. MS medium supplemented with NAA showed the highest germination rate (76.67%) and optimal growth, producing 3.33 shoots per explant, 8 cm shoot length, 9.66 cm root length, and 9.33 lateral roots. Media supplemented with 2,4-D had a 66.67% germination rate, 3 shoots per explant, 8.33 cm shoot length, and 25 lateral roots. BAP resulted in moderate shoot formation (1.33 shoots per explant) with no lateral root induction. Control explants produced a single shoot. For micropropagation, the control and BAP at 0.5 mg/L resulted in optimal microcutting success rate (55.2%), with 2.06 shoots per explant, 3.34 cm shoot length, and 1 leaves. Higher concentrations of BAP (1.0 and 1.5 mg/L) reduced shoot and leaf development. The combination of 1.5 mg/L BAP with 1 mg/L NAA increased the microcutting success rate to 66.7%, although shoot length (0.86 cm) and leaf count (1.53) were reduced. This study is limited to in vitro conditions and a specific range of PGRs concentrations. Further research is needed to explore environmental factors in ex vitro culture. These findings provide an efficient protocol for enhancing Asparagus propagation, offering practical benefits for horticultural production, conservation, and pharmaceutical applications. This study presents original data on the effect of specific PGRs on both germination and micropropagation, revealing the potential of NAA, 2,4-D and BAP treatments in optimizing A. officinalis development under controlled conditions.