Two-dimensional (2D) molybdenum disulfide (MoS2) with a unique action on H2was incorporated into 1,2-bis(triethoxysilyl)ethane (BTESE)-derived bridged microporous organosilica networks to form a composite membrane for H2separation by a sol–gel method. Due to their opposite ζ-potentials, a continuous surface without lamellar boundary defects was formed between BTESE sols derived by the hydrolysis–polymerization reaction and MoS2nanosheets. When the MoS2content increased in BTESE networks, the H2permeance showed an overall increasing trend in the range of 1.85–2.89 × 10–7mol·m–2s–1Pa–1(552–864 GPU), which was higher than that of pristine BTESE membrane with the H2permeance of 491 GPU. In addition, optimized MoS2/BTESE membranes showed a much higher H2/N2permselectivity of 129 than that of the pristine BTESE membrane of 17 at 100 °C. The synergistic effect of BTESE and MoS2nanosheets plays an important role. Through adsorption isotherm test and diffusivity as well as energy calculation, BTESE networks became denser by nonporous MoS2addition that prevented N2from passing, while H2was promoted with excellent adsorption on charged edges of MoS2, resulting in improved H2separation performance both in permeance and selectivity. This provides an attractive mechanism for hydrogen separation.