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A general review of the sub-seafloor biosphere is presented. This includes an update and assessment of prokaryotic cell distributions within marine sediments, current deepest 1922 m, and the impact of this on global sub-seafloor biomass estimates. These global estimates appear relatively robust to different calculation approaches and our updated estimate is 5.39 × 10²⁹ cells, taking into consideration new data from very low organic matter South Pacific Gyre sediments. This is higher than other recent estimates, which is justified as several sediments, such as gas hydrate deposits and oil reservoirs, can have elevated cell concentrations. The proposed relationship between elevated cell concentrations and Milankovitch Cycles in sequential diatom rich layers at some sites, demonstrates not only a dynamic deep biosphere, but also that the deep biosphere is an integral part of Earth System Processes over …