Gamma-amino-butyric acid receptors (GABAARs) and glycine receptors (GlyRs) are the principal inhibitory neurotransmitter receptors in the mammalian brain and for GABAARs, in particular, represent a key pharmacological target for a range of CNS disorders. In order to determine the pharmacological properties and composition of GABAARs and GlyRs expressed in human excitatory cortical neurones (hECNs) derived from human pluripotent stem cells (see Bilican et al., 2014; Livesey et al., 2014), we have evaluated the effects of an array of subunit-selective pharmacological ligands on whole-cell GABAARs-mediated and GlyR-mediated currents recorded from hECNs. For GABAARs, the agonists GABA and muscimol showed relatively low potencies with mean values (± standard error from fit) for their EC50s of 256 ± 9 μM (n=12) and 182 ± 10 μM (n=6), respectively. GABAAR-mediated currents were blocked by the antagonists bicuculline and picrotoxin with IC50 values of 2.7 ± 0.7 µM (n=5) and 5.2 ± 0.2 µM (n=4), respectively. GABAARs were determined to be predominantly γ-subunit containing as indicated by the sensitivity of GABA-evoked currents to diazepam and Zn2+, together with the observation that gaboxadol showed only weak agonist action. Potentiation of currents by propofol and etomidate, and lack of current inhibition by salicyline salycylhydrazide, indicated that GABAARs in hECNs contain either the β2 or β3-subunit. In addition, our study suggests that the principal α-subunit in GABAARs present in hECNs is likely to be the α3-subunit; this conclusion was also supported by Western blot analysis of GABAAR subunit expression. Thus, the properties of expressed GABAARs are consistent with a native, immature cortical phenotype and appear to be comprised of α3, β2/3 and γ subunits. For GlyRs, the glycine EC50 was determined as 167 ± 20 µM (n=7) while the IC50 values for picrotoxin and strychnine were 191 ± 16 µM (n=5) and 708 ± 64 nM (n=3). These results suggest hECNs express GlyR population likely containing a mixture of both homomeric and heteromeric receptors comprised of α1, α2, and β subunits, a finding that was also confirmed by RT-PCR analysis. For both GABAARs and GlyRs our data suggest the presence of receptor populations that are consistent with those expressed in late embryonic and early postnatal cortical neurons.