Plants have long been recognized as vital sources of a myriad of chemically, structurally, and functionally diversified compounds, utilized by humankind since ancient times. Phytoconstituents, often termed metabolites, are referred as primary and secondary metabolites. Primary metabolites constitute basic compounds involved in growth, development, and primary functions. Secondary metabolites, derived from primary metabolic pathways, serve secondary functions such as pollination, defence, and stress management. These secondary metabolites find multiple uses in diverse industries, with the majority employed in medicine. However, a major limitation lies in the quantities in which they are produced. Certain compounds are produced in highly specialized systems, leading to limitations such as organ specificity and season dependence, hindering large-scale production and commercial utilization. In this context, plant tissue culture offers numerous options for in vitro production of secondary metabolites. Plant cell, tissue and organ culture primarily utilized for large scale propagation of elite genotypes, however utilized to produce phytochemicals. This chapter provides an overview of in vitro secondary metabolite production, with an emphasis on the different strategies employed in plant biotechnology.