Use of Human Primary Stem Cell Cultures for Screening Cell Proliferati on Sti mulatory and Inhibitory Eff ects of Botanical Preparati ons: an Alternati ve to Animal Models

Abstract

Animals are widely used in scienti fi c research as highly specifi c ‘models’ of humans. Primary cell culture is a widely used model to reduce and replace the use of animal models, as per the 3R concept. We investi gated cell sti mulatory eff ects of selected botanical preparati ons on primary human stem cell cultures, in place of animal models. Primary human fi broblast stem cell (hFSC), human mesenchymal stem cell (hMSC), and human haematopoieti c stem cell (hHSC) cultures, were established in-house, and characterized by immunophenotyping. Varying concentrati ons of selected botanical preparati ons (mature leaf concentrate of Carica papaya Sri Lankan wild type culti var [MLCC], disti llates of Vernoina/ Mallotus [VMD], and Ficus benghalensis [FBD]) were tested ex vivo for cell proliferati on sti mulati on on these cell platf orms, using the MTT assay. Compared with untreated controls, signifi cant proliferati ve eff ects were demonstrated by MLCC on female (at 0.2% concentrati on) and male (0.6%) hMSCs, and on male and female hHSCs (0.6-0.2%) (P<0.05). FBD (0.05-1.2%) sti mulated signifi cant proliferati on of both male hMSCs, and male hHSCs, but inhibited proliferati on of female hMSCs (P<0.05), with no signifi cant eff ects on female hHSCs. Conversely, VMD (0.2-1%) showed signifi cant inhibitory eff ects on both male and female hMSCs, yet at 0.6% showed signifi cant proliferati ve eff ects on female hHSCs (P<0.05). Selected VMD concentrati on exhibited approximately 1.5-folds higher % cell sti mulati on than the positi ve control (p < 0.01). Thus, eff ecti ve cell proliferati ve concentrati ons of botanical preparati ons were identi fi ed using primary stem cell lines, aligning with “replacement” as per the 3R concept.