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Isolation, purification and characteristics of mononuclear phagocytes from human placentas
Abstract:
Methods for the isolation of mononuclear phagocytes from human placentas by digestion with collagenase or with trypsin are described. Mononuclear phagocytes were approximately 35% of the cells obtained. Preparations were enriched for mononuclear phagocytes by sequential density gradient centrifugation over Ficoll-Hypaque and Percoll, adherence to plastic and removal of contaminating trophoblastic cells with brief trypsin exposure. Monolayers obtained by these methods were greater than 85% mononuclear phagocytes. These cells were predominantly of fetal origin; most were mature macrophages but up to 20% appeared to be recently derived from blood monocytes as determined by ultrastructural peroxidase cytochemistry.
Insights
Researchers developed methods to isolate mononuclear phagocytes from human placentas. These placental cells, primarily macrophages, offer a valuable resource for studying immune cell development and function.
Area of Science:
- Immunology
- Cell Biology
- Reproductive Biology
Background:
- Human placenta is a rich source of immune cells.
- Mononuclear phagocytes play crucial roles in fetal-placental interactions and immune regulation.
Purpose of the Study:
- To describe effective methods for isolating mononuclear phagocytes from human placentas.
- To characterize the isolated cells for purity and origin.
Main Methods:
- Enzymatic digestion of placental tissue using collagenase or trypsin.
- Sequential density gradient centrifugation (Ficoll-Hypaque and Percoll).
- Adherence to plastic and selective removal of trophoblastic cells.
Main Results:
- Initial digests yielded approximately 35% mononuclear phagocytes.
- Enrichment methods resulted in >85% pure mononuclear phagocyte monolayers.
- Isolated cells were predominantly of fetal origin, comprising mature macrophages and recently derived monocytes.
Conclusions:
- Established protocols allow for efficient isolation of highly purified mononuclear phagocytes from human placenta.
- These placental-derived mononuclear phagocytes represent a valuable model for studying macrophage and monocyte biology in a pregnancy context.