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Quantitative analysis of Weibel-Palade bodies
Alice Liu1,2, Christopher J Ng2
1Department of Bioengineering, Washington University, St. Louis, MO, United States of America.
Plos One
|December 21, 2022
Summary
Confluency significantly impacts Weibel-Palade body (WPB) generation in endothelial cells. However, the method used to achieve cell confluency does not affect WPB quantification, offering flexibility in cell culture techniques.
Area of Science:
- Endothelial cell biology
- Cellular vesicle formation
- Biotechnology and cell culture
Background:
- Weibel-Palade bodies (WPBs) are crucial endothelial cell vesicles storing von Willebrand factor (VWF).
- Cellular confluency is known to influence WPB numbers, but optimal culture methods require clarification.
Purpose of the Study:
- To compare two distinct cell culture methods for achieving endothelial cell confluency.
- To determine the impact of these methods on Weibel-Palade body (WPB) quantification.
- To assess the role of confluency in WPB generation in different endothelial cell types.
Main Methods:
- Tested two confluency methods: constant incubation (96 hrs) with varying seeding density, and constant seeding density (30,000 cells/cm²) with varying incubation time.
- Utilized human umbilical vein endothelial cells (HUVECs) and endothelial colony-forming cells (ECFCs).
- Quantified nuclei count, total WPB count, and WPB/nuclei ratio for each condition.
Main Results:
- Both increasing incubation time and seeding density showed a trend towards increased nuclei and WPB counts.
- No significant difference in WPB/nuclei ratio was observed between the two confluency methods.
- Confluent cultures exhibited significantly higher WPB/nuclei counts compared to sub-confluent cultures.
Conclusions:
- Cellular confluency is a major determinant of WPB generation in endothelial cells.
- The choice between constant incubation time or constant seeding density does not influence WPB quantification.
- These findings provide valuable insights into optimizing cell culture for WPB analysis.
