Adaptive Metal Ion Transport and Metalloregulation Drive Fate Differentiation in Pluripotent Synthetic Cells

Abstract

Abstract Pluripotent stem cells can yield different cell types depending on a sequence of differentiation signals as it activates/deactivates functions and keeps a memory of previous inputs. Herein, we achieve pluripotency in synthetic cells with three dormant apo-metalloenzymes such that they can differentiate towards different fates depending on the sequence of specific metal ion transport with ionophores. In the first differentiation step, the selective transport of extracellular metal ion cofactors into pluripotent giant unilamellar vesicles (GUVs) differentially activates enzymatic pathways that give rise to an increase of intracellular pH, production of hydrogen peroxide, or cell lysis. Formerly added ionophores suppress transport with subsequent ionophores due to in membrane interactions between ionophores. Consequently, the addition of a second ionophore leads to a dampened response in the multipotent GUV and a third ionophore in no further response, reminiscent of a terminally differentiated GUV. Taken together, the pluripotent GUV differentiates into five final fates depending on the sequence of three ionophores by virtue of adaptive metal ion transport.