There are many types of non-metallic MgAl2O4 inclusions observed in Al-deoxidized steel coupling with Mg treatment, including single-particle MgAl2O4, agglomerated MgAl2O4, and MgAl2O4-MnS. Thermodynamic calculation shows that MgAl2O4 precipitates in the liquid phase. The phase transformation follows liquid + Al2O3 + MgAl2O4 → liquid + MgAl2O4 → liquid + MgO + MgAl2O4 → liquid + MgO with the Mg content increasing when the Al content is a constant in molten steel, and it is in agreement with the experimental results for the formation of MgAl2O4 in molten steel. The calculation results of various attractive forces between two particles show that the cavity bridge force plays a dominant role in the agglomeration process and results in the agglomerated MgAl2O4. The lattice mismatch calculation result shows that MgAl2O4 can provide effective sites for MnS nucleating in steel.