Identification of compounds that enhance differentiation of progenitor cells constitutes an important goal in regenerative medicine. Herein we sought to test the efficacy of extracts from Tridax procumbens (TP - a medicinal plant rich in secondary metabolites with ethnopharmacological activities) on the differentiation of cultured embryonic mouse pancreas and liver. TP leaves were authenticated, air-dried, blended, soaked in ethanol, and concentrated to obtain the extract. The extract was fractionated using Hexane, Dichloromethane, and Ethyl Acetate to obtain Fractions (HF, DMF, and EAF, respectively). E11.5-day explants (pancreas, liver) were isolated, cultured for five days, maintained in a humidified incubator, and treated with (20 g/ml) dimethyl sulfoxide (control), extract or fractions for 48 hours. The expression of pancreatic endocrine (insulin, glucagon), exocrine (amylase), ductal (cytokeratin-7, peanut agglutinin), mesenchymal (vimentin) cells, and the liver (alpha-fetoprotein) protein markers were evaluated using immunofluorescence detection. Confocal images were collected (n≥3) and processed with Image J and Zen 2.3 lite software. Data were analyzed using ANOVA and student’s t-test, with p &lt; 0.05 considered as significant. The DMF-treated hepatoblasts elicited a 1.5-fold increase in alpha-fetoprotein expression compared to the control. Enhanced branching morphogenesis of the ductal system was observed in DMF-treated cultured embryonic pancreas based on increased expression of cytokeratin 7 and peanut agglutinin. The DMF-treated pancreata showed enhanced expression of insulin (β-cells), vimentin (mesenchymal cells) and amylase (acinar cells) (all 1.3-fold increased); and glucagon (α-cells, 2.8-fold increased), relative to control. TP did not have any adverse effects in terms of cell death in either tissue explant. In conclusion, TP induced differentiation of pancreatic endocrine and acinar cell types and enhanced the population of AFP-expressing hepatoblasts (bipotent liver precursors that generate hepatocytes and ductal cells). Identifying the active component(s) of the extracts and elucidating the mechanism of action is an important future goal.

Semi-purified fraction of Tridax procumbens induces differentiation of mouse embryonic pancreas and enhances the population of liver progenitor cells

ACS Spring 2022

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