Hematopoietic stem cells (HSCs) are continuously stimulated by physical interactions with bone marrow or umbilical cord niches as well as by chemical factors found within these niches. The niche can be mimicked by modification of the cytokine composition, elasticity, topography, and/or charge. This work employed cell culture plates coated with several concentrations of poly-L-lysine (PLL), a positively charged synthetic amino-acid chain. Culture substrates that employed relatively high initial coating concentrations of PLL significantly increased the total number of HSCs during ex vivo expansion of CD34(+) cells, as well as erythroid differentiation. Furthermore, the 0.01% PLL substrate stimulated enucleation of erythroid cells, leaving behind a number of extruded nuclei at the bottom of the culture plate, followed by an increase in the number of erythrocytes. Thus, PLL will likely prove useful to enhance the expansion of HSCs and erythroid cells, in addition to the generation of red blood cells.