Fig. 1

Flowchart of experimental mouse BPD model. C57/BL6 mice were time-mated and on day 16 of pregnancy (E16) injected with either 0.1 μg lipopolysaccharide (LPS) in 5 μl saline or an equal volume of vehicle into each amniotic sac. Pups were then allowed to deliver naturally at term. a Determine the optimal dosage. Pups were placed either in normoxia or hyperoxia chambers on PND3.5 (12 hours before hAEC treatment). On PND4, either hAECs (50,000, 75,000 or 100,000 cells) or saline were injected intravenously through the superficial temporal vein using the automated microinjection system as per the intra-amniotic injections. Mouse pups were culled on PND7. b Determine the optimal route of administration. Pups were exposed to either normoxia or hyperoxia from PND3.5, then the optimal dose of hAECs were injected intratracheally on PND4 and compared to the intravenous injection groups in the dose-effect experiment. The pups were culled on PND14. c Assess timing of administration. Pups were exposed to either normoxia or hyperoxia on PND0, and intravenous administration of 100,000 hAECs was given either on PND0.5 or on PND4. Pups were culled either on PND7 or PND14. d Long-term study. Pups were given hAECs intravenously either on PND0.5 or PND4, and were kept in the hyperoxia chamber from PND0 to PND28, when they were weaned and transferred to standard housing facilities. Animals were culled at either 6 or 10 weeks of age. hAEC human amnion epithelial cell, IT intratracheal, IV intravenous