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Isolation of villous microvessels from the human placenta
Abstract:
A method for isolating the microvessels of the human placental villi has been developed in order to culture perivascular cells. It consists of an initial selection of the villi by serial sieving. The villi retained by the 75 microns sieve were digested by collagenase-dispase. A Percoll gradient permitted the isolation of microvessels still surrounded by stromal fibres and cells. Another digestion by collagenase-dispase eliminated the contaminant elements and allowed, after a new Percoll gradient, microvessels with endothelium, basement membrane and a few perivascular cells to be obtained. Each step of the isolation of microvessels was monitored by light or electron microscopy. Our study confirms the isolation of microvessels embedded in their basement membrane and the preservation of endothelial and perivascular cells after digestion. This method, which has permitted the culture of placental endothelial cells and pericytes, appears of interest for studying microvascular angiogenesis and permeability.
Insights
Researchers developed a new method to isolate human placental microvessels for culturing perivascular cells. This technique enables the study of microvascular angiogenesis and permeability.
Area of Science:
- Vascular Biology
- Cell Biology
- Human Placental Research
Background:
- Human placental villi contain microvessels crucial for nutrient exchange.
- Previous methods for isolating placental microvessels have limitations for cell culture.
- Perivascular cells and endothelial cells are important for vascular function.
Purpose of the Study:
- To develop and validate a novel method for isolating intact human placental microvessels.
- To enable the subsequent culture of endothelial cells and perivascular cells from isolated microvessels.
- To provide a tool for studying microvascular angiogenesis and permeability in the human placenta.
Main Methods:
- Serial sieving of human placental villi to select specific size fractions.
- Enzymatic digestion using collagenase-dispase to release microvessels.
- Density gradient centrifugation with Percoll to isolate microvessels.
- Microscopic evaluation (light and electron) to monitor isolation steps.
Main Results:
- Successful isolation of human placental microvessels retaining their basement membrane.
- Preservation of endothelial cells and associated perivascular cells after isolation.
- Demonstration of microvessels suitable for subsequent cell culture.
Conclusions:
- The developed method effectively isolates intact human placental microvessels.
- This technique supports the isolation and culture of placental endothelial cells and pericytes.
- The method is valuable for research into microvascular angiogenesis and permeability.