We give a comprehensive study from flavor observables of pion, kaon, D_(s), and B_(s) mesons for limiting the Two Higgs Doublet Models (2HDMs) with natural flavor conservation, namely, Z_2 symmetric and aligned type of models. With use of the updated studies and analyses of B -> tau nu, D -> mu nu, D_s -> tau nu, D_s -> mu nu, K -> mu nu, Pi -> mu nu, B^0_s -> mu^+ mu^-, B^0_d -> mu^+ mu^-, tau -> K nu, tau -> Pi nu, B -> X_s gamma, K-K bar mixing, B^0_d-B^0_d bar mixing, and B^0_s-B^0_s bar mixing, we obtain constraints on the parameters in the 2HDMs. To calculate the constraints, we pay attention to a determination of CKM matrix elements and re-fit them to experimental data so that new contributions from additional Higgs bosons do not affect the determination. In addition, we discuss excesses of observables in the muon anomalous magnetic moment and the semi-tauonic B meson decays in the context of the 2HDM.