An aeroponic seed potato production technology project has been established in Pakistan through the Technical Cooperation Project (TCP) between RDA-KOPIA and PARC. For sustainable potato production, producing high-quality and disease-free potato seeds is critical in Pakistan. The majority of the farmers recycle their own seeds or obtain them through informal ways. Consequently, tuber-borne illnesses proliferate, seeds deteriorate, and yield diminishes. To mitigate such problems, the country moved to sustainable seed potato production by adopting rapid seed multiplication strategies like the combination of tissue culturing with aeroponic seed production technology which is being evaluated as an alternative to traditional pre-basic seed production methods. A series of experiments were carried out to evaluate the shoot development and mini tuber production attributes between two cultivars, different plantlet size and planting period between October 27, 2021, and January 27, 2022, at the NARC aeroponic facility. A CRD design with three replications was used. The study aimed to compare the productivity by variety, plantlet size and planting period to improve aeroponic seed production technology in Pakistan. The results indicated that Lady Rosetta outperformed Asterix in terms of shoot growth and small tuber output. Early planting at an optimal temperature of 27 degrees Celsius in the daytime showed a substantial difference when compared to late planting in November and tuberization was commenced 50-55 days after transplantation. Furthermore, in an aeroponic seed potato production system, normal plantlets (8-10 cm) at the time of transplantation, performed better than medium and small plantlets. In a nutshell, it was determined that normal size plantlets (8-10 cm) of Lady Rosetta cultivar, planted at optimum time to plant showed best results in the aeroponic system at NARC, Islamabad, Pakistan.