In vitro culture has become a key tool to produce seed potato (Solanum tuberosum L.) tubers, a crop of great global importance. This systematic review, based on the PRISMA-ScR methodology, analyzes the main biotechnological strategies used to obtain high-quality, healthy, and pathogen-free seeds, overcoming the limitations of traditional propagation methods. A comprehensive search was conducted in Scopus, Web of Science, and ScienceDirect (June 2025), prioritizing 65 experimental studies published between 2010 and 2025 in indexed journals. Techniques such as meristem culture, micropropagation, microtuber production, the use of temporary immersion bioreactor systems (TIBs), and synthetic seed generation were examined. These methodologies offer advantages such as accelerated propagation, higher yields, reduced use of agrochemicals, germplasm conservation, and economic efficiency. TIBs stand out for improving the survival and productivity of basic seed. This review is organized around four axes: applied techniques, key procedures, economic impact and sustainability, and future perspectives. This work constitutes a useful guide for optimizing seed tuber production using plant biotechnology. Potato, with an annual production of 400 million tons, is positioned as one of the most important food crops globally, comparable in importance to wheat, rice, and maize [ 2 ]. Indeed, its high nutritional value makes it the third or fourth most important food crop worldwide [ 3 , 4 ]. Given its importance in global food security [ 5 ], the production of high-quality seed tubers using advanced techniques, such as in vitro culture, is essential to meet the growing demand [ 6 ]. This technique offers significant advantages over traditional methods by allowing rapid and efficient propagation of disease-free plants, which is crucial for ensuring crop quality and yield [ 7 ]. Microtubers Culture The production of in vitro tubers, or “microtubers”, emerged as an efficient alternative for the propagation and storage of germplasm, with key protocols being established in the late 1970s and early 1980s [ 31 ]. Microtubers culture allows the production of small tubers under controlled conditions, ideal for the propagation of crops such as potatoes and their efficient storage. These microtubers, generated in vitro from plant tissues, have similar characteristics to field tubers and can be directly planted [ 10 , 11 ]. Their implementation has proven to be an efficient alternative, with significant improvements in crop survival and yield compared to other propagation methods [ 1 , 12 ]. 3.4. Propagation in Bioreactors The use of bioreactors for micropropagation in liquid media marked a significant advance towards automation, being key to the adaptation of temporary immersion bioreactor systems (TIBs) in mass production [ 34 ]. Bioreactors are highly efficient systems for the mass propagation of potato plants and microtubers. In this sense, (TIBs) stand out as an element of special relevance, given their capacity to optimize plant development through periodic immersions in liquid media [ 8 , 35 ]. In addition, it is critical that mother plants exhibit high yield and superior tuber quality, ensuring that these characteristics are passed on to propagated plants [ 26 , 46 ]. Phytosanitary quality: It is imperative to use plant material free of viruses, bacteria, and fungi to prevent contamination and guarantee the healthiness of in vitro cultures. In the field of agriculture, the use of tissue culture techniques, such as thermotherapy, chemotherapy, and electrotherapy, has proven to be effective in the production of pathogen-free planting material. In addition, these procedures have been shown to contribute to the preservation of germplasm and the development of transgenic plants [ 21 ]. They are also used in the process of seed multiplication and germplasm distribution, thus ensuring crop health [ 10 ]. The process of microtuber production under in vitro conditions is characterized by the presence of two main phases. First, plant proliferation is carried out, in which explants generate new shoots through optimized culture conditions, promoting their vegetative growth. In the field planting process, these larger tubers tend to give rise to plants with a greater number of stems and a greater number of Sanitation is an important aspect of seed tuber production, as historically, it has lacked strict regulations, which can make crops vulnerable to viral and bacterial diseases, as well as to pest attacks [ 10 ]. The implementation of biotechnology in the production chain, such as micropropagation, guarantees propagation material with high phytosanitary quality [ 26 ]. In particular, in vitro culture using apical meristems is essential for establishing a healthy variety bank and guaranteeing the absence of viral infections in plants [ 35 ]. In regions such as the Andes, where traditional production often lacks regulations and standardized techniques, vulnerability to viral and bacterial diseases, as well as pests, generates cost overruns, yield losses, and a decrease in crop quality [ 10 , 45 ]. The native potato, an underutilized tuber, is exposed to a high risk of spreading pathologies through the tubers used for planting, resulting in reduced agricultural yields [ 45 ]. In contrast, in vitro production of potato seed tubers with high genetic and phytosanitary standards emerges as an economically viable option [ 10 ]. Soilless technologies, such as aeroponics and hydroponics, have proven effective in improving crop health and optimizing seed tuber production. Simultaneously, germplasm conservation through cryopreservation protects potato genetic diversity, while biotechnology contributes to crop improvement, strengthening food security, and agricultural efficiency [ 29 ]. Despite the progress made in the field of potato micropropagation, there are still areas that require further exploration and rigorous study.