Computational model and microfluidic platform for the investigation of paracrine and autocrine signaling in mouse embryonic stem cells

Autocrine and paracrine signaling mechanisms are traditionally difficult to study due to the recursive nature of the process and the sub-micromolar concentrations involved. This has proven to be especially limiting in the study of embryonic stem cells that might rely on such signaling for viability, self-renewal, and proliferation. To better characterize possible effects of autocrine and paracrine signaling in the setting of expanding stem cells, we developed a computational model assuming a critical need for cell-secreted survival factors. This model suggested that the precise way in which the removal of putative survival factors could affect stem cell survival in culture. We experimentally tested the predictions in mouse embryonic stem cells by taking advantage of a novel microfluidic device allowing removal of the cell-conditioned medium at defined time intervals. Experimental results in both serum-containing and defined N2B27 media confirmed computational model predictions, suggested existence of unknown survival factors with distinct rates of diffusion, and revealed an adaptive/selective phase in mouse embryonic stem cell response to a lack of paracrine signaling. We suggest that the described computational/experimental platform can be used to identify and study specific factors and pathways involved in a wide variety of paracrine signaling systems.