Harvest time is one of the key factors for obtaining high-quality cherry tomatoes. This parameter depends on environmental conditions and tomato variety. In plant factories with artificial lighting (PFALs), it is possible to control environmental conditions to enhance tomato production and quality. Since the ripening status of tomato fruit is correlated with cumulative temperature (CT), and the temperature inside PFALs can be easily controlled, CT could be used as an alternative method to predict tomato harvest time. In this study, three experiments were conducted to determine the optimal CT for harvesting high-quality cherry tomatoes in a PFAL. The experiments aimed to (1) evaluate the yield and quality of cherry tomatoes as affected by different harvest times based on CT (ranging from 900 to 1400 °C), (2) comparatively evaluate the yield and quality of cherry tomatoes that were still on the plant and off the plant (in storage) based on the same CT levels (i.e., 1100, 1200, and 1300 °C), and (3) investigate the fruit-cracking percentage during the ripening stage based on CT levels. The results showed that the fruit harvested at lower CTs exhibited higher hardness values, while those harvested at higher CTs had a higher sugar content. The on-the-plant treatment resulted in a higher yield and sugar content compared with the off-the-plant treatment, indicating that harvesting tomatoes early would come at the expense of a certain yield and sweetness. Moreover, the fruit-cracking percentage tended to increase with increasing CT, possibly due to the fast fruit growth rate and increased internal turgor pressure. These results indicated that producers can use CT as an index to predict the harvest time, thereby optimizing profits in cherry tomato production.