Paeonia lactiflora Pall. (herbaceous peony) is a high-value ornamental and medicinal plant in China with considerable market potential. However, conventional propagation methods are limited by low multiplication rates and long production cycles, making it difficult to meet the demand for large-scale planting materials. As a key approach for rapid propagation, tissue culture techniques for P. lactiflora have achieved significant progress in recent years. This review summarizes advances in the tissue culture system for P. lactiflora over the past decade, focusing on major in vitro regeneration pathways (organogenesis and somatic embryogenesis) and crucial technical stages, including explant selection and culture environment optimization. Distinct from previous reviews that only introduce partial technical aspects of P. lactiflora tissue culture, this review comprehensively outlines the overall tissue culture system, analyses the current species-specific bottlenecks (browning, vitrification, rooting and acclimatization) with their underlying causes and proposes targeted strategies. Furthermore, future development trends are prospected by integrating emerging research directions, including molecular regulatory mechanisms and eco-adaptive breeding. This review aims to provide a theoretical foundation and technical support for obtaining propagule for commercial plantations and achieving multi-functional utilization of P. lactiflora.