Mechanical stimuli can be introduced to three dimensional (3D) cell cultures by use of perfusion bioreactor. Especially in musculoskeletal tissues, shear stress caused by fluid flow generally increase extra-cellular matrix (ECM) production and cell proliferation. The relationship between the shear stress and the tissue development in situ is complicated because of the non-uniform pore distribution within the cell-seeded scaffold. In this study, we firstly demonstrated that Doppler optical coherence tomography (DOCT) is capable of monitoring localized fluid flow and shear stress in the complex porous scaffold by examining their variation trends at perfusion rate of 5, 8, 10 and 12 ml/hr. Then, we developed the 3D porous cellular constructs, cell-seeded chitosan scaffolds monitored during several days by DOCT. The fiber based fourier domain DOCT employed a 1300 nm superluminescent diode with a bandwidth of 52 nm and a xyz resolution of 20×20×15 μm in free space. This setup allowed us not only to assess the cell growth and ECM deposition by observing their different scattering behaviors but also to further investigate how the cell attachment and ECM production has the effect on the flow shear stress and the relationship between flow rate and shear stress in the developing tissue construct. The possibility to monitor continuously the constructs under perfusion will easily indicate the effect of flow rate or shear stress on the cell viability and cell proliferation, and then discriminate the perfusion parameters affecting the pre-tissue formation rate growth.