Lang Bian ginseng (Panax vietnamensis var. langbianensis) is a rare and traditional herbal medicine with many pharmacological activities. It was first discovered by Duy et al. (2016) in Lam Dong province, Vietnam. Until now, there has been only one publication on Lang Bian ginseng micropropagation via somatic embryogenesis (Anh et al., 2022). However, this study still had limitations, including the small number of somatic embryos and the time-consuming. This study investigated the effect of spermidine, glutamine, and proline on the enhancement of somatic embryogenesis (SE) of Lang Bian ginseng using thin cell layer culture technology. The results showed that the optimal SE rate and the number of somatic embryos per explant were achieved on MS medium supplemented with 0.01 mM spermidine in leaf explants (93.32 % and 54.20 embryos, respectively) and petiole explants (96.66 % and 68.80 embryos, respectively) and higher compared to these in other treatments, including control, glutamine, and proline treatments. Besides, adding 0.01 mM spermidine to the culture medium also improved the quality of somatic embryos, mainly cotyledonary stage, through an enhanced synthesis of antioxidant enzymes [catalase (CAT) and superoxide dismutase (SOD), and ascorbate peroxidase (APX)]. In addition, the fluctuations of endogenous hormones during initiation and maturation of SE derived from 0.01 mM spermidine treatment were recorded. Endogenous CKs (ZEA, 2iP, KIN, mT), IAA, and GA3 concentrations were the highest during the induction stage in both explants. Meanwhile, the remaining endogenous hormones (MEL, ABA, and SA) exhibited no inevitable fluctuation trends. In addition, IAA was only detected at the induction stage of SE in both explants. Moreover, cotyledonary somatic embryos derived from 0.01 mM spermidine treatment grew well in MS medium supplemented with 1.2 mg/L AgNPs.