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 WJMSCS 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 ß-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.

Occurrence of vertebrate steroids, estradiol 17beta and progesterone in the reproducing females of the mud crab Scylla serrata.

In crustaceans, vitellogenesis is known to be controlled by eyestalk neuropeptides, biogenic amines, ecdysteroids and a juvenile hormone-like compound, methyl farnesoate. In recent years, the occurrence of vertebrate steroid hormones, estradiol 17beta (E2) and progesterone (PG) has also been reported in a few decapods, although their precise role in female reproduction is yet to be determined. The levels of E2 and PG in the ovary, hepatopancreas and the hemolymph of the red mud crab, Scylla serrata were analyzed in different vitellogenic stages in order to establish a correlation between hormone profile and stages of vitellogenesis. It was observed that the levels of both the steroids increased steeply in the tissues at the onset of vitellogenesis (vitellogenic stage I). Maximum levels of estradiol were present in the hepatopancreas whereas the highest concentration of progesterone was seen in the ovary, suggesting dichotomous roles for these hormones in vitellogenesis. Furthermore, levels of these hormones were estimated in different embryonic stages of the eggs of the sand crab Emerita asiatica and mud crab S. serrata. Their levels fluctuated, following a definite pattern in the different stages, suggesting a possible functional role as morphogenetic hormones. This study, in addition, also reports the presence of E2 and PG on lipovitellin purified from ovary and eggs as well as vitellogenin purified from the hemolymph implicating a role for these lipoproteins as steroid carriers.