Pluripotin stimulates the growth of hematopoietic stem cells while suppressing the expansion of bone marrow mesenchymal stem cells and fibroblasts

Abstract

Hematopoietic stem cells (HSCs) are essential in the production and maintenance of red blood and immune cells. Small molecules that target HSC modulators may aid in the proliferation and expansion of HSCs. To that end, we investigated the effect of two small molecules on HSC expansion: pluripotin (an ERK1 and RasGAP inhibitor) and CHIR-99021 (a GSK-3 inhibitor). After 7 days of treatment, both Pluripotin and CHIR-99021 resulted in a 3-fold increase in the murine pool of HSCs in a dose-dependent manner. Furthermore, we looked into the effect of Pluripotin on the ex vivo expansion of human umbilical cord blood and bone marrow mononuclear cells. Pluripotin treatment, in particular, increased human CD34+ and ALDHbr HSC content up to threefold when compared to the control. Furthermore, Pluripotin treatment increased the number of human CD133+ HSC cells by a factor of five. Intriguingly, Pluripotin treatment reduces bone marrow-derived mesenchymal stem cell (MSC) proliferation and fibroblast growth while having no effect on adipose-derived MSCs. CHIR-99021 treatment had no effect on MSC or fibroblast proliferation. In conclusion, pluripotin-induced stem cell expansion is unique to HSCs and can be used to expand HSCs while suppressing unwanted fibroblast or MSC growth in primary ex vivo cultures.