A complete protocol of Nolana intonsa I. M. Johnst, micropropagation was developed. The culture media used were WPM, DKW, and MS as control. DKW medium produced the best results over in vitro development of N. intonsa, generating a significant increase in the multiplication rate, lateral shoots formation, height, and general quality of the plantlets. In the case of fresh weight, dry weight, and water content, no differences were observed between MS and DKW, but a significant decrease in these parameters was produced when the WPM medium was used. Subsequently, when DKW was defined as basal medium, the effect of auxin supplementation was also evaluated. The addition of IBA generated a desirable effect over the in vitro plantlets. The addition of 0.05 mg L-1 of IBA improve the multiplication rate, the shoot's development, plant's height, and reduced in a significant way the damage to the plantlets. Fresh weight, dry weight, and water contents also showed a significant increase when IBA supplementation was added to the DKW medium. The plants where acclimatization of the plantlets was possible, obtaining finished pot plants capable of growth under greenhouse conditions. The development of a micropropagation protocol of N. intonsa could help the conservation and sustainable utilization of this endemic Chilean species.