Polymesoda arctata is a Caribbean estuarine clam commercially important that currently is considered threatened. The reproductive conditioning, spawning and early development were evaluated under laboratory conditions using organisms collected in the lagoon system of Ciénaga Grande de Santa Marta (Colombian Caribbean) between December 2020 and May 2021. Two reproductive conditioning experiments were conducted over 7 weeks, examining the frequency of mature animals in the natural environment (controls) and in the laboratory under two water temperatures (26 and 29 °C) and different salinity regimes (constant at 5 and 15 ppt, increasing from 5 to 15 ppt, and decreasing from 15 to 5 ppt). One spawning experiment was done, comparing the spawning response of adult clams exposed to various stimuli (changes in water temperature and salinity, flow of seawater irradiated with UV light, addition of a high concentration of food and sperm to the water, as well as injections of serotonin and KCl into the foot and/or adductor muscle). Higher frequencies of mature clams were observed after 2 to 5 weeks of reproductive conditioning at a low temperature (26 °C) and under a constant salinity regime (at 5 or 15 ppt). Higher percentages of spawning and post-stimulation survival as well as spawning response speed were observed in specimens injected in the foot with a low concentration of serotonin (0.4 mL at 20 μM). The time of spawning response onset was shorter in males than in females. The fecundity increased with the live weight of the females, showing an average value of 3.4 × 106 oocytes per female. The early development of the estuarine clam from oocyte to juvenile stage was described in detail. The growth rates of larvae and spat were relatively high but their survival values were low, showing the need for additional research on the larval and post-larval rearing.