Human Primary Stem Cell Cultures as Screening Platforms for Botanical Preparations as Alternatives to Animal Models: Evaluating Cell Proliferation Stimulatory and Inhibitory Effects

Animals are widely used as human “models” in scientific research, yet primary human stem cell cultures provide an alternative under the 3R (Reduction, Replacement and Refinement) concept to reduce and replace animal models. This study investigated the cell stimulatory effects of selected botanical preparations on primary human stem cell cultures. In-house established primary human fibroblast (hFSC), mesenchymal (hMSC), and haematopoietic (hHSC) stem cells, derived from surgical waste, were characterized by immunophenotyping. The proliferation effects of Carica papaya leaf concentrate (MLCC), Vernonia/Mallotus (VMD), and Ficus benghalensis (FBD) distillates were assessed using the MTT assay. MLCC significantly stimulated proliferation in male and female hMSCs and hHSCs (p<0.05). FBD promoted proliferation in male hMSCs and hHSCs but inhibited female hMSCs. VMD inhibited both male and female hMSCs yet stimulated female hHSCs at 0.6%. Notably, VMD at selected concentrations exhibited approximately 1.5-fold higher cell stimulation than the positive control (p<0.05). These findings highlight effective botanical concentrations for cell proliferation, demonstrating the utility of primary human stem cell platforms as “models” in biomedical research. This approach aligns with the “replacement” principle of the 3R concept, promoting ethical and human-relevant alternatives to animal testing.