The derivation of regionally-defined cellular phenotypes from human pluripotent stem cells (hPSCs) presents a platform for the assessment of human physiology and disease in vitro. We have developed a protocol that generates enriched populations of oligodendrocyte (OL)-lineage cells from multiple hPSC lines. Within 1 week of OL differentiation, approximately 10-20% of the cells in culture were found to be platelet-derived growth factor receptor α (PDGFRα)-positive OL precursor cells (OPCs) and between 60-80% were OLs that were positive for mature marker, myelin basic protein. The membrane properties of rodent OPCs and OLs are well described and demonstrate distinctive intrinsic membrane properties and ion channel expression. In this regard, we have examined the membrane properties of hPSC-derived OPCs and followed these through to their maturation into OLs. Depolarization of OPCs induced large, sustained outwardly rectifying currents that were substantially reduced in OLs. Our data indicate a change in the ion channel expression profile of OLs compared to those expressed in OPCs. Passive membrane properties were measured as indicators of development. For OPCs compared to OLs input resistances of 2786 ± 228 MΩ (OPC, n = 11) vs 1508 ± 122 MΩ (OL, n = 12), whole-cell capacitances of 5.7 ± 0.5 pF (OPC, n = 11) vs 29.6 ± 3.7 pF (OL, n = 11), and resting membrane potentials of -34 ± 1 mV (OPC, n = 11) vs -44 ± 1.3 mV (OL, n = 12) indicate maturation of OPCs into OLs. We next examined the properties of AMPA receptors (AMPARs) expressed in hPSC-derived OPCs and OLs. AMPARs can be composed of 4 potential subunits, GluA1-4 of which the GluA2 subunit is developmentally post-transcriptionally edited post-transcriptionally, such that an ion channel-lining glutamine (Q) is RNA-edited to arginine (R). This imparts distinctive biophysical and pharmacological properties to AMPARs which include low Ca2+-permeability and reduced single-channel conductance. Using non-stationary fluctuation analysis we found that the single-channel conductance of AMPARs expressed in OPCs was 8.2 ± 0.9 pS (n = 7) and in OLs was 3.2 ± 0.5 pS (n = 7). This is consistent with a developmental switch in AMPAR composition from GluA2(R)-lacking to GluA2(R)-containing AMPARs in OPCs and OLs, respectively. This was confirmed by assessing block of AMPAR-mediated currents by NASPM which inhibited currents in OPCs 54 ± 9.8% ( n = 8) but by only 28.1 ± 5.6% in OLs(n = 7). Thus, ion channel expression profiles exhibited by hPSC-derived OPCs and OLs are in good agreement with native OL-lineage cell development seen in rodents and offer the potential to assess the physiological properties of OL-lineage cells derived from patients suffering from demyelinating diseases.