Experimental evidence suggesting a type of glucose uptake regulation prevailing in resting and differentiated cells was surveyed. This type of regulation is characterized by transport-limited glucose metabolism and depends on segregation of glucose transporters on microvilli of differentiated or resting cells. Earlier studies on glucose transport regulation and a recently presented general concept of influx regulation for ions and metabolic substrates via microvillar structures provide the basic framework for this theory. According to this concept, glucose uptake via transporters on microvilli is regulated by changes in the structural organization of the microfilament bundle, which is acting as a diffusion barrier between the microvillar tip compartment and the cytoplasm. Both microvilli formation and the switch of glucose metabolism from "metabolic regulation" to "transport limitation" occur during differentiation. The formation of microvillar cell surfaces creates the essential preconditions to establish the characteristic functions of specialized tissue cells including the coordination between glycolysis and oxidative phosphorylation, regulation of cellular functions by external signals, and Ca²⁺ signaling. The proposed concept integrates various aspects of glucose uptake regulation into a ubiquitous cellular mechanism involved in regulation of transmembrane ion and substrate fluxes.