In South Africa, more than 30 small, enclosed game reserves have reintroduced lions over the last two decades, which now house more than 500 individuals. There is a high risk of inbreeding in these fragmented, fenced and isolated populations, which may be compounded by a lack of management guidelines. A population of 11 founder lions Panthera leo was reintroduced to Madikwe Game Reserve in 1995, and this population has in turn become a source for reestablishing other populations. Only four lineages were reintroduced, founder males were related to founder females, and since 1997, only one male lineage maintained tenure for >9 years, resulting in breeding with direct relatives. Interventionist management to limit lion population growth and inbreeding in Madikwe has taken the form of translocating, trophy hunting and culling of mainly sub adult lions. Despite this management, inbreeding started 5 years after reintroduction. Reproductive performance and thus population growth in Madikwe were dependent on the overall lion population density. When lion density was low, females first gave birth at a significantly younger age and produced larger litters, resulting in a high population growth rate, which decreased significantly when lion density in the park reached carrying capacity, that is, 61 lions. This might have profound consequences for future reestablishment of lion populations when restocking new reserves: our study illustrates the need for founder populations of reintroduced endangered predator species to be as large and genetically diverse as possible, and thereafter new genetic material should be supplemented. The development of such management guidelines is becoming very important as large predator populations become increasingly fragmented and managed as metapopulations.