Utilizing PCL microcarriers for high-purity isolation of primary endothelial cells for tissue engineering

Guang-Zhen Jin, Jeong-Hui Park, Eun-Jung Lee, Ivan B Wall, Hae-Won Kim

Research output: Contribution to journalArticlepeer-review

Abstract

Endothelial cells (ECs) are widely used in research, both for fundamental vascular biology research and for exploring strategies to create engineered vascularized tissues. Primary isolation often results in contamination from fibroblasts and vascular smooth muscle cells that can potentially affect function, particularly during the initial expansion period needed to establish the cell culture. In the current study, we explored the use of microcarriers to selectively isolate ECs from the lumen of intact vessels to enhance the purity during the isolation procedure. First, rat aortic explant culture was performed and after 2 weeks of culture, flow cytometry revealed that only 60% of the expanded cell population was positive for the endothelial marker CD31. Then, we employed a strategy to selectively isolate ECs and improve their purity by introducing microcarriers to the lumen of intact aorta. After 10 days, microcarriers were carefully removed and placed in cell culture dishes and at 15 days, a large near confluent layer of primary ECs populated the dish. Flow cytometry revealed that >90% of the expanded cells expressed CD31. Moreover, the cells were capable of forming tubule-like structures when plated onto Matrigel, confirming their function also. The highly modular and transportable nature of microcarriers has significant potential for isolating ECs at high purity, with minimal contamination.

Original languageEnglish
Pages (from-to)761-768
Number of pages8
JournalTissue engineering. Part C, Methods
Volume20
Issue number9
Early online date16 Jun 2014
DOIs
Publication statusPublished - Sep 2014

Keywords

  • Animals
  • Aorta/cytology
  • Cell Proliferation/drug effects
  • Cell Separation/methods
  • Cell Shape/drug effects
  • Cells, Cultured
  • Endothelial Cells/cytology
  • Fluorescent Antibody Technique
  • Male
  • Organ Culture Techniques
  • Phenotype
  • Polyesters/pharmacology
  • Rats, Sprague-Dawley
  • Tissue Engineering/methods
  • Tissue Scaffolds/chemistry

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