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Artificial Perfusion of the Fetal Circulation of the In situ Mouse Placenta: Methodology and Validation

By Helen Bond, B. Baker, R.D.H. Boyd, E. Cowley, J.D. Glazier, C.J.P. Jones, Colin P. Sibley, B.S. Ward and S.M. Husain


Here we present methodology and validation (including measurement of unidirectional maternofetal clearance (Kmf) of 45Ca and 14C-mannitol) for in situ perfusion of the mouse placenta. On day 18 of gestation (term = 19 days) mice were anaesthetised and the uterus delivered into a saline bath (40°C). A fetus was selected, the umbilical artery and vein catheterised and perfused with Krebs Ringer (pH 7.4) at 60 μl/min. 45Ca/14C-mannitol (2 μCi/5 μCi in 50 μl saline) was injected via maternal tail vein. Perfusate samples were collected every 5 min for 45 min. Maternal carotid artery pressure was monitored throughout perfusion. A terminal maternal cardiac blood sample was taken and analysed. Placentas were immersion fixed and processed for electron microscopy. Kmf for 45Ca and 14C-mannitol was calculated as perfusate [45Ca or 14C-mannitol] × perfusion rate/maternal plasma [45Ca or 14C-mannitol] × placental weight. Maternal cardiac blood chemistry at termination (n = 8–15, mean ± SEM) was as follows: pH 7.153 ± 0.016, PCO2 45.48 ± 2.06 mmHg, PO2 66.47 ± 7.10 mmHg, Na+ 151.4 ± 1.2 mmol/l, K+ 5.54 ± 0.17 mmol/l, Ca2+ 1.15 ± 0.03 mmol/l, glucose 7.2 ± 0.5 mmol/l, and lactate 1.76 ± 0.77 mmol/l. A successful 45 min perfusion in which perfusate recovery was >95% occurred in >50% of animals. Perfusion did not alter placental morphology or carotid pressure. Kmf (μl/min/g placenta) for 45Ca (66.0 ± 8.4 (n = 7)) was significantly higher than Kmf for 14C-mannitol (20.0 ± 2.4 (n = 5)) (p < 0.01). These data demonstrate physiological perfusion of the mouse placenta in situ and its usefulness for measurement of solute transfer

Topics: QH301
Publisher: Elsevier
Year: 2006
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