Establishment and characterization of conditionally immortalized endothelial cell lines from the thoracic duct and inferior vena cava of tsA58/EGFP double-transgenic rats

Mitsuhiro Matsuo, Keiichi Koizumi, Sanae Yamada, Masatoshi Tomi, Ri Ichi Takahashi, Masatsugu Ueda, Tetsuya Terasaki, Masuo Obinata, Ken Ichi Hosoya, Osamu Ohtani, Ikuo Saiki

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22 Citations (Scopus)

Abstract

The basic biology of blood vascular endothelial cells has been well documented. However, little is known about that of lymphatic endothelial cells, despite their importance under normal and pathological conditions. The lack of a lymphatic endothelial cell line has hampered progress in this field. The objective of this study has been to establish and characterize lymphatic and venous endothelial cell lines derived from newly developed tsA58/EGFP transgenic rats harboring the temperature-sensitive simian virus 40 (SV40) large T-antigen and enhanced green fluorescent protein (EGFP). Endothelial cells were isolated from the transgenic rats by intraluminal enzymatic digestion. The cloned cell lines were named TR-LE (temperature-sensitive rat lymphatic endothelial cells from thoracic duct) and TR-BE (temperature-sensitive rat blood-vessel endothelial cells from inferior vena cava), respectively, and cultured on fibronectin-coated dishes in HuMedia-EG2 supplemented with 20% fetal bovine serum and Endothelial Mitogen at a permissive temperature, 33°C. A temperature shift to 37°C resulted in a decrease in proliferation with degradation of the large T-antigen and cleavage of poly (ADP-ribose) polymerase. TR-LE cells expressed lymphatic endothelial markers VEGFR-3 (vascular endothelial growth factor receptor), LYVE-1 (a lymphatic endothelial receptor), Prox-1 (a homeobox gene product), and podoplanin (a glomerular podocyte membrane mucoprotein), together with endothelial markers CD31, Tie-2, and VEGFR-2, whereas TR-BE cells expressed CD31, Tie-2, and VEGFR-2, but no lymphatic endothelial markers. Thus, these conditionally immortalized and EGFP-expressing lymphatic and vascular endothelial cell lines might represent an important tool for the study of endothelial cell functions in vitro.

Original languageEnglish
Pages (from-to)749-758
Number of pages10
JournalCell and Tissue Research
Volume326
Issue number3
DOIs
Publication statusPublished - 2006 Dec 1

Keywords

  • Endothelial cell
  • Enhanced green fluorescent protein
  • Inferior vena cava
  • Rat (tsA58/EGFP double-transgenic)
  • Temperature-sensitive large T-antigen
  • Thoracic duct

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

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    Matsuo, M., Koizumi, K., Yamada, S., Tomi, M., Takahashi, R. I., Ueda, M., Terasaki, T., Obinata, M., Hosoya, K. I., Ohtani, O., & Saiki, I. (2006). Establishment and characterization of conditionally immortalized endothelial cell lines from the thoracic duct and inferior vena cava of tsA58/EGFP double-transgenic rats. Cell and Tissue Research, 326(3), 749-758. https://doi.org/10.1007/s00441-006-0229-x