14-Deoxy-11,12-didehydroandrographolide Promotes Ex Vivo Expansion of Umbilical Cord Blood Stem Cells through Stemness-Related Gene Regulation

Abstract

Umbilical cord blood (UCB) is a valuable alternative source of hematopoietic stem and progenitor cells (HSPCs) for allogeneic transplantation. However, the limited number of HSPCs that can be obtained from a single UCB unit remains a significant clinical challenge. Therefore, strategies to enhance the ex vivo expansion of functional HSPCs are of considerable interest. A key requirement for improving the success of HSC-based therapies, including transplantation and gene editing, is the ability to expand or preserve functional human HSCs during ex vivo culture. In this study, we demonstrate that 14-deoxy-11,12-didehydroandrographolide (14-DDA), a diterpenoid isolated from Andrographis paniculata, significantly promotes the ex vivo expansion of UCB-derived CD34⁺ cells and enriches for primitive HSPC subsets (CD34⁺CD38^–CD90⁺). Functional assays reveal that 14-DDA enhances colony-forming unit output, which preserves multilineage differentiation potential. Furthermore, 14-DDA activates the Wnt/β-catenin signaling pathway, indicating a potential mechanism for supporting stem cell self-renewal. Gene expression profiling reveals upregulation of stemness- and proliferation-associated genes, while suppression of genes is related to differentiation, stress responses, and oncogenesis. These findings suggest that 14-DDA promotes the expansion of functional HSPCs while maintaining their primitive phenotype and reducing oncogenic risk, highlighting its potential as a natural small molecule for enhancing hematopoietic stem cell-based therapies.