Background: Healthcare workers (HCW) are at high risk to develop mental health problems during the COVID-19 pandemic because of additional work load, perceived stress, and exposure to patients with COVID-19. Currently, there are few studies on change over time in the prevalence of depressive symptoms during pandemic start among HCW. Thus, the aims of the current study were to examine whether depressive symptoms increased during the pandemic and were associated with perceived stress and own COVID-19 infection and workplace exposure to virus-infected patients. Methods: The cohort study used longitudinal data from HCW collected monthly (July 2020 till December 2020) during the first year of the pandemic before vaccination became available. The sample of n = 166 was drawn from a German hospital and included medical (e.g. nurses, therapists, and physicians) and administrative staff. Using multilevel models, we analyzed the change in depressive symptoms [assessed with General Depression Scale (GDS), a validated German version of the Center for Epidemiological Studies Depression Scale (CES-D)] and its association with perceived stress across the study period. Laboratory-confirmed own infection was tested as a potential moderator in this context. Subscales of the GDS were used to examine change over time of depressive symptom modalities (e.g. emotional, somatic, and social interactions (β, 95% confidence interval). Results: Depression scores increased significantly during the study period (β = .03, 95% CI [0.02, 0.05]). Perceived stress was associated with depressive symptoms (β = .12, 95% CI [0.10, 0.14]) but did not change over time. Exposure to COVID-19 infection was associated with a higher increase of depressive symptoms (β = .12, 95% CI [0.10, .14]). Somatic symptoms of depression increased among medical HCW with workplace exposure to COVID-19 (β = .25, 95% CI [0.13, 0.38]), but not in administrators (β = .03, 95% CI [−0.04, 0.11]). Conclusion: Research is needed to identify factors that promote the reduction of depressive symptoms in medical HCW with exposition to COVID-19 patients. Awareness of infection protection measures should be increased.