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Purification and characterization of rat bone marrow endothelial cells
Experimental Hematology
|November 1, 1986
Summary
Researchers developed a method to isolate pure, viable endothelial cells from rat bone marrow using Percoll gradient centrifugation and elutriation. This technique yields high-purity marrow endothelium for in vitro research.
Area of Science:
- Cell Biology
- Hematology
- Biotechnology
Background:
- Endothelial cells (ECs) are crucial for vascular function.
- Obtaining pure, viable ECs from bone marrow is challenging.
- Marrow-derived ECs are valuable for research.
Purpose of the Study:
- To establish a reliable method for isolating high-purity, high-viability endothelial cells from rat bone marrow.
- To enable in vitro studies using enriched populations of marrow ECs.
Main Methods:
- Bone marrow cell suspensions were prepared using collagenase.
- Discontinuous gradient centrifugation on Percoll (1.04 and 1.06 densities) concentrated ECs.
- Centrifugal elutriation further purified the endothelial cell fraction.
Main Results:
- Electron microscopy confirmed endothelium purity ranging from 63% to 90%.
- Flow cytometry and fluorescent microscopy showed 49%-51% factor VIII-positive cells.
- Cell viability exceeded 90% with a yield of approximately 10^6 cells.
Conclusions:
- A robust method for isolating rat bone marrow endothelial cells was developed.
- The method ensures high purity and viability for experimental applications.
- This technique supports in vitro research on marrow endothelium.