Duckweeds can be potentially used in human and animal nutrition, biotechnology or wastewater treatment. To cultivate large quantities of a defined product quality, a standardized production process is needed. A small-scale, re-circulating indoor vertical farm (IVF) with artificial lighting and a nutrient control and dosing system was used for this purpose. The influence of different light intensities (50, 100 and 150 µmol m−2 s−1) and spectral distributions (red/blue ratios: 70/30, 50/50 and 30/70%) on relative growth rate (RGR), crude protein content (CPC), relative protein yield (RPY) and chlorophyll a of the duckweed species Lemna minor and Wolffiella hyalina were investigated. Increasing light intensity increased RGR (by 67% and 76%) and RPY (by 50% and 89%) and decreased chlorophyll a (by 27% and 32%) for L. minor and W. hyalina, respectively. The spectral distributions had no significant impact on any investigated parameter. Wolffiella hyalina achieved higher values in all investigated parameters compared to L. minor. This investigation proved the successful cultivation of duckweed in a small-scale, re-circulating IVF with artificial lighting.