<p>Due to numerous problems in production practice, such as diseases and pests, long planting period, autotoxicity, and decreasing genetic purity of germplasm, this study introduces a tissue culture regeneration system for the medicinal plant <i>Codonopsis pilosula</i>. This system has increased the production efficiency of <i>Codonopsis pilosula</i> by more than 50%. Root, stem, and leaf of <i>Codonopsis pilosula</i> as explants were cultured in medium with suitable PGRs to induce callus, multiple shoots, and roots, eventually resulting in the regeneration of whole plants by somatic embryogenesis and organogenesis. For somatic embryogenesis, the root serves as the optimal explant for callus induction, with a combination 0.5&#xa0;mg·L⁻¹ 2,4-D, 1.0&#xa0;mg·L⁻¹ 6-BA, and 0.5&#xa0;mg·L⁻¹ KT being the most suitable for generating callus. The callus generated from the root is embryonic and can grow into An embryoid. In addition, 0.5&#xa0;mg·L⁻¹ 6-BA and 0.5&#xa0;mg·L⁻¹ NAA significantly improve the induction of shoots regenerated from callus. For organogenesis, the optimal combination for multiple shoots growth is 1.5&#xa0;mg·L⁻¹ 6-BA and 0.5&#xa0;mg·L⁻¹ NAA, for example the stem with a pair of leaves as explant Also, 0.2&#xa0;mg·L⁻¹ IBA significantly improves the rooting of <i>Codonopsis pilosula</i>. The ISSR analysis indicates there are no changes in the genomic DNA of the regeneration plantlets, signifying that the regenerated plantlets exhibit high genetic stability. After callus induction, it takes less than 4 mo to obtain regenerated plantlets that are genetically stable, grow healthily, and have high antioxidant activity.</p>