Bamboo forests, due to rapid growth and short harvest rotation, play an important role in carbon cycling and local economic development. Accurate estimation of bamboo forest aboveground biomass (AGB) has garnered increasing attention during the past two decades. However, remote sensing-based AGB estimation for bamboo forests is challenging due to poor understanding of the mechanisms between bamboo forest growth characteristics and remote sensing data. The objective of this research is to examine the remote sensing characteristics of on-year and off-year bamboo forests at different dates and their AGB estimation performance. This research used multiple Sentinel-2 data to explore AGB estimation of bamboo forests in Zhejiang Province, China, by taking into account the unique characteristics of on-year and off-year bamboo forest growth features. Combining field survey data and Sentinel-2 spectral responses (spectral bands and vegetation indices) and textural images, random forest was used to identify key variables for AGB estimation. The results show that (1) the on-year and off-year bamboo forests have considerably different spectral signatures, especially in the wavelengths between red edge 2 and near-infrared wavelength (NIR2) (740–865 nm), making it possible to separate on-year and off-year bamboo forests; (2) on-year bamboo forests have similar spectral signatures although AGB increases from as small as 40 Mgha−1 to as high as 90 Mgha−1, implying that optical sensor data cannot effectively model on-year bamboo AGB; (3) off-year bamboo AGB has significant relationships with red and shortwave infrared (SWIR) spectral bands in the April image and with red edge 2 in the July image, but the AGB saturation problem yields poor estimation accuracy; (4) stratification considerably improved off-year bamboo AGB estimation but not on-year, non-stratification using the April image is recommended; and (5) Sentinel-2 data cannot solve the bamboo AGB data saturation problem when AGB is greater than 70 Mgha−1, similar to other optical sensor data such as Landsat. More research should be conducted in the future to integrate multiple sources—remotely sensed data (e.g., lidar, optical sensor data) and ancillary data (e.g., soil, topography)—into AGB modeling to improve the estimation. The use of very high spatial resolution images that can effectively extract tree density information may improve bamboo AGB estimation and yield new insights.