A synthetic formulation, Dhanwantharam kashaya, delays senescence in stem cells

Objectives: Mesenchymal stem cells (MSCs) derived from post-natal tissues offer a suitable source of MSCs for cellular therapy. Limitation of the use of MSCs for therapeutic purposes is attributed to the onset of senescence and slowing down of proliferation upon repeated passaging. Dhanwantram kashaya (DK), a synthetic herbal formulation, is widely used in Ayurvedic medicine as a growth stimulant in children and for nerve regeneration. In this study, we evaluated the effects of DK on the proliferation, viability and senescence of human Wharton jelly MSCs (WJMSCs) in vitro. Results: Using the MTT proliferation assay and live/dead trypan blue analysis, we found that DK increased proliferation of WJMSC_S up to three folds when supplemented in the culture media. The BrdU cell proliferation assay showed a substantial increase in WJMSCs treated with DK. Notably, the β-galactosidase senescence assay revealed that drug treated WJMSCs at late passage still had intact and viable WJMSCs whereas the untreated cells exhibited profound senescence. Conclusion: These studies indicate that DK enhances the quality of WJMSCs by not only increasing the proliferation rate and decreasing their turnover time but also by delaying senescence. We have, thus, identified for the first time that a traditional Ayurvedic formulation, Dhanwantram kashaya, used as a growth enhancer, is able to improve the yield and quality of stem cells in vitro and could be an effective non-toxic supplement for culturing WJMSCs for clinical applications.