Isolation of macrophages (Hofbauer cells) from human term placenta and their prostaglandin E2 and thromboxane production

Placental macrophages (Hofbauer cells) are located close to trophoblast cells and fetal capillaries, which makes them ideal candidates for involvement in regulatory processes within the villous core. Their production of various cytokines and prostaglandin (PG) synthesizing enzymes has previously bee...

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Bibliographic Details
Published in:Human reproduction (Oxford) 1997-04, Vol.12 (4), p.847-852
Main Authors: Wetzka, B, Clark, D E, Charnock-Jones, D S, Zahradnik, H P, Smith, S K
Format: Article
Language:English
Subjects:
Biological and medical sciences
Cell Separation
Chorionic Villi - metabolism
Delivery, Obstetric
Dinoprostone - biosynthesis
Female
Fundamental and applied biological sciences. Psychology
Humans
Immunoenzyme Techniques
Immunohistochemistry
Macrophages - metabolism
Mother. Fetoplacental unit. Mammary gland. Milk
Placenta - cytology
Placenta - metabolism
Pregnancy
Pregnancy. Parturition. Lactation
Thromboxane A2 - biosynthesis
Vertebrates: reproduction
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Summary:Placental macrophages (Hofbauer cells) are located close to trophoblast cells and fetal capillaries, which makes them ideal candidates for involvement in regulatory processes within the villous core. Their production of various cytokines and prostaglandin (PG) synthesizing enzymes has previously been shown immunohistochemically. Hofbauer cells were isolated from human placenta after term deliveries by Ficoll and Percoll gradient centrifugation. Remaining trophoblast cells were removed with anti-epidermal growth factor (EGF)-receptor-coated Dynabeads followed by differential adherence. The identity of isolated cells was investigated by immunohistochemistry with anti-CD68, which showed that >90% cells were positive. After a 36 h recovery period in either 20% O2 or 5% O2, fresh medium was applied and PGE2 and thromboxane (TXA2) production analysed by enzyme immunoassay at 4, 8, and 24 h. PGE2 and TXA2 were both produced by placental macrophages with PGE2 synthesis being predominant. Concentrations of both could be stimulated by lipopolysaccharide with maximum effect after 24 h. Culture in low oxygen caused decreased PGE2 concentrations, whereas TXA2 production remained unchanged. In conclusion, the presented isolation protocol allows further study of Hofbauer cell function. This study also presents novel findings regarding the prostaglandin production of term Hofbauer cells under normal and hypoxic conditions.
ISSN:0268-1161
1460-2350
DOI:10.1093/humrep/12.4.847