Artificial Perfusion of the Fetal Circulation of the In situ Mouse Placenta: Methodology and Validation
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)....
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Veröffentlicht in: | Placenta (Eastbourne) 2006-04, Vol.27, p.69-75 |
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Sprache: | eng |
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Zusammenfassung: | 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,
PCO
2 45.48
±
2.06
mmHg,
PO
2 66.47
±
7.10
mmHg, Na
+ 151.4
±
1.2
mmol/l, K
+ 5.54
±
0.17
mmol/l, Ca
2+ 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. |
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ISSN: | 0143-4004 1532-3102 |
DOI: | 10.1016/j.placenta.2006.01.005 |