Titanium-based alloys are susceptible to hydrogen embrittlement (HE), a phenomenon that deteriorates fatigue properties. Ti-6Al-4V is the most widely used titanium alloy and the effect of hydrogen embrittlement on fatigue crack growth (FCG) was investigated by carrying out crack propagation tests in air and high-pressure H2 environment. The FCG test in hydrogen environment resulted in a drastic increase in crack growth rate at a certain Δ K, with crack propagation rates up to 13 times higher than those observed in air. Possible reasons for such behavior were discussed in this paper. The relationship between FCG results in high-pressure H2 environment and microstructure was investigated by comparison with already published results of cast and forged Ti-6Al-4V. Coarser microstructure was found to be more sensitive to HE. Moreover, the electron beam melting (EBM) materials experienced a crack growth acceleration in-between that of cast and wrought Ti-6Al-4V.