Pharmacokinetic parameter estimation with the support of dynamic diffuse fluorescence tomography (DFT) can provide helpful diagnostic information for tumor differentiation and monitoring. Adaptive extended Kalman filtering (AEKF) as a nonlinear filter method has the merits of high quantitativeness, noise robustness, and initialization independence. In this paper, indirect and direct AEKF schemes combining with a commonly used two-compartment model were studied to estimate the pharmacokinetic parameters based on our self-designed dynamic DFT system. To comprehensively compare the performances of both schemes, the selection of optimal noise covariance matrices affecting estimation results was first studied, then a series of numerical simulations with the metabolic time ranged from 4.16 min to 38 min was carried out and quantitatively evaluated. The comparison results show that the direct AEKF outperforms the indirect EKF in estimation accuracy at different metabolic velocity and demonstrates stronger stability at the large metabolic velocity. Furtherly, the in vivo experiment was conducted to achieve the indocyanine green pharmacokinetic-rate images in the mouse liver. The experimental results confirmed the capability of both schemes to estimate the pharmacokinetic-rate images and were in agreement with the theory predictions and the numerical simulation results.