Plasmids are extrachromosomal DNA elements of microorganisms encoding beneficial genetic information. They were thought to be equally distributed to daughter cells during cell division. Here we use mathematical modeling to investigate the evolutionary stability of plasmid segregation for high-copy plasmidsplasmids that are present in up to several hundred copies per cellcarrying antibiotic resistance genes. Evolutionary stable strategies (ESS) are determined by numerical analysis of a plasmid-load structured population model. The theory predicts that the evolutionary stable segregation strategy of a cell depends on the plasmid copy number: For low and medium plasmid load, both daughters receive in average an equal share of plasmids, while in case of high plasmid load, one daughter obtains distinctively and systematically more plasmids. These findings are in good agreement with recent experimental results. We discuss the interpretation and practical consequences.Author summary In the last years, it becomes more and more clear that heterogeneity in isogenic bacterial populations is rather the rule than the exception. This observation is interesting as it reveals the complex social life of bacteria, and also because of tremendous practical implications in medicine, biotechnology, and ecology. The central questions in this field are the identification of the underlying proximate causes (molecular mechanisms) on the one hand and on the other hand the identification of ultimate causes (evolutionary forces) that shape the social life of bacteria. We focus on plasmid dynamics, in particular on plasmid segregation. Recent experiments showed that plasmid segregation depends on the plasmid load. We identify possible evolutionary factors that shaped this process. It turns out that the ambivalence in the effect of plasmidsadvantageous if present in low copy numbers, a metabolic burden if present in high copy numbersis able to explain the experimental observations. The experimental findings can be interpreted as a variant of the principle of division of labor, as it is well known from e.g. persister cells or sporulation. Our model extends the theory of unequal segregation of damage to the ambivalent role of plasmids. Similarly, it is known that certain gene regulatory proteins are acting in a dose-dependent manner. Due to differences in their cellular concentrations and in their affinities to various target promoters, differential gene expression patterns are achieved. Consequently, tight concentration control is observed . Another example is the strict copy-dependent utilization of autolysins during cell division. These enzymes carefully open the bacterial cell wall to allow for its extension . Overproduction of these enzymes leads to cell lysis . These molecules are in principle also candidates for a segregation characteristic similar to that of high copy plasmids described here.