Alpine plants represent a unique group of high-altitude flora that thrive under extreme environmental conditions categorized by low temperature, high ultraviolet radiation, reduced atmospheric pressure and short growing seasons. These adaptive species produce a diverse array of bioactive secondary metabolites with significant pharmaceutical value, supporting traditional and modern medicine systems. However, habitat loss, climate change and unsustainable harvesting have placed many Himalayan alpine species under severe threat, making conservation and sustainable utilization an urgent priority. Conventional propagation approaches are often constrained by seed dormancy, low germination rates and recalcitrant tissues, necessitating alternative biotechnological interventions. In vitro techniques including micropropagation, callus-mediated regeneration, somatic-embryogenesis, cryopreservation and production of synthetic seeds, have emerged as reliable strategies for mass multiplication, germplasm conservation and metabolite enhancement of critically endangered alpine taxa. This chapter includes advancements made in key Himalayan alpine plants such as Aconitum spp., Angelica spp., Arnebia spp., Fritillaria spp., Picrorhiza spp., Podophyllum spp., Rhodiola spp., Saussurea spp., Swertia spp., Trillium spp. and Valeriana spp. The evaluation of tissue culture studies highlights optimization of explant choice, hormonal combinations, regeneration pathways and conservation protocols. The compiled knowledge not only underscores the potential of plant tissue culture for overcoming recalcitrance and ensuring ex situ conservation but also provides a framework for future research on biotechnological interventions to safeguard alpine biodiversity and promote sustainable utilization of these valuable resources.