OBJECTIVES: The purpose of this study was to evaluate cell proliferation in platelet-enriched collagen plugs with and without addition of amniotic fluid-derived heterologous foetal cells to seal an iatrogenic membrane defect in the foetal rabbit model. METHODS: Amniotic fluid cells were harvested from three donor does at 23 days of gestation (term = 32 days) and labelled with carboxyfluorescein diacetate succinimidyl ester (CFDA-SE). In 42 other does, foetal membrane defects were induced by foetoscopic needle puncture at 23 days of gestation, and closed with either a platelet-enriched collagen plug with (n = 44) or without (n = 32) amniotic fluid cells. At 30 days of gestation, the defects were harvested and assessed microscopically. RESULTS: The plugs enriched with heterologous amniotic fluid cells more commonly had proliferating cells in the centre of the plug than those without cell addition. CFDA-SE labelling confirmed the presence of heterologous amniotic fluid cells over the entire membrane plug. Cell typing showed a mixture of fibroblasts and epithelial cells at the wound edges, whereas in the centre, there was an abundance of fibroblasts. CONCLUSION: When sealing iatrogenic membrane defects in the foetal rabbit model, enrichment of collagen plugs with platelets and amniotic fluid-derived heterologous foetal cells increases local cell proliferation.