Fatigue crack growth (FCG) tests were conducted on a medium-Mn steel annealed at two intercritical annealing temperatures, resulting in different austenite (γ) to ferrite (α) phase fractions and different γ (meta-)stabilities. Novel in-situ hydrogen plasma charging was combined with in-situ cyclic loading in an environmental scanning electron microscope (ESEM). The in-situ hydrogen plasma charging increased the fatigue crack growth rate (FCGR) by up to two times in comparison with the reference tests in vacuum. Fractographic investigations showed a brittle-like crack growth or boundary cracking manner in the hydrogen environment while a ductile transgranular manner in vacuum. For both materials, the plastic deformation zone showed a reduced size along the hydrogen-influenced fracture path in comparison with that in vacuum. The difference in the hydrogen-assisted FCG of the medium-Mn steel with different microstructures was explained in terms of phase fraction, phase stability, yielding strength and hydrogen distribution. This refined study can help to understand the FCG mechanism without or with hydrogen under in-situ hydrogen charging conditions and can provide some insights from the applications point of view.