Soil organic matter (SOM) plays an important role in the global carbon cycle, especially in alpine ecosystems. However, ongoing forest expansion in high-elevation systems potentially alters SOM storage through changes in organic matter (OM) inputs and microclimate. In this study, we investigated the effects of an Picea abies L. afforestation chrono-sequence (0 to 130 years) of a former subalpine pasture in Switzerland on soil organic carbon (SOC) stocks and SOM dynamics. We found that SOC stocks remained constant throughout the chrono-sequence, with comparable SOC stocks in the mineral soils after afforestation and previous pasture (SOC forest40 = 11.6 ± 1.1 kg m−2, SOC forest130 = 11.0 ± 0.3 kg m−2 and SOC pasture = 11.5 ± 0.5 kg m−2). However, including the additional carbon of the organic horizons in the forest, reaching up to 1.7 kg m−2 in the 55-year old forest, resulted in an increase in the overall SOC stocks following afforestation. We found that the soil C:N ratio in the mineral soil increased in the topsoil (0–5 cm) with increasing forest stand age, from 11.9 ± 1.3 in the pasture to 14.3 ± 1.8 in the 130-year old forest. In turn, we observed a decrease in the soil C:N ratio with increasing depth in all forest stand ages. This suggests that litter-derived organic matter (C:N from 35.1 ± 1.9 to 42.4 ± 10.8) is likely to be incorporated and translocated from the organic horizon to the mineral topsoil (0–10 cm) of the profiles. Due to the high root C:N ratio (pasture 63.5 ± 2.8 and forests between 54.7 ± 3.9 and 61.2 ± 2.9), particulate root-derived organic matter seems to have a rather small effect on forest soil C:N ratios, as well as on SOC accumulation in the mineral soil. These results suggest that, although afforestation does not change the SOC stock in the mineral soil, there is an apparent alteration in the SOM dynamics through changes in the litter composition caused by the vegetation shift. We conclude that, at our study site, spruce afforestation on a former subalpine pasture does not change the total SOC stock and that, consequently, there is no additional SOC sequestration on a decadal to centennial scale.