Fresh rocks sampled from the 14.0°S hydrothermal field of the South Atlantic Ridge can be divided into two categories: olivine-gabbro and basalt. The olivine-gabbro is composed mainly of three types of minerals: olivine, clinopyroxene and plagioclase, while a multitude of melt inclusions occur in the plagioclase phenocrysts of the basalts. We analyzed the whole-rock, major and trace elements contents of the basalts, the mineral chemistry of phenocrysts and melt inclusions in the basalts, and the mineral chemistry of olivine-clinopyroxene-plagioclase in the olivine-gabbro, then simulated magma evolution within the crust using the COMAGMAT program. The whole-rock geochemistry shows that all the basalts exhibit typical N-MORB characteristics. In addition, the mineral chemistry characteristics of the olivine-gabbro (low-Fo olivine, low-Mg# clinopyroxene, high-TiO₂ clinopyroxene, low-An plagioclase), show that strong magma differentiation occurred within the crust. Nevertheless, significant discrepancies between those minerals and phenocrysts in the basalts (high-Fo olivine, high-An plagioclase) reflect the heterogeneity of magma differentiation. High Mg# (≈0.72) melt inclusions isobaric partial crystallization simulations suggest that the magma differentiation occurred at the depth shallower than 13.03 km below the seafloor, and both the vertical differentiation column shows distinct discrepancies from that of a steady-state magma chamber. Instead, a series of independent magma intrusions probably occurred within the crust, and their corresponding crystallized bodies, as the primary high-temperature thermal anomalies within the off-axis crust, probably act as the heat source for the development of the 14.0°S hydrothermal system.