Establishment of bone marrow-derived endothelial cell lines from ts-SV40 T-antigen gene transgenic rats

K. Hattori, M. Muta, M. Toi, H. Iizasa, M. Shinsei, T. Terasaki, M. Obinata, M. Ueda, E. Nakashima

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21 Citations (Scopus)

Abstract

Purpose. Postneonatal neovascularization is thought to result exclusively from the proliferation, migration, and remodeling of fully differentiated endothelial cells (ECs). Recently, it has been reported that bone marrow contains cells which can differentiate into ECs and contribute to neoangiogenesis in adult species. In this study, we tried to establish conditionally immortalized endothelial cell lines (TR-BME) derived from rat bone marrow. Methods. Mononuclear cells were isolated and differentiated into ECs at 37°C from the bone marrow of a transgenic rat harboring temperature-sensitive SV40 large T-antigen (ts T-Ag) gene. Then, the cells were transferred and incubated at 33°C, a permissive temperature for ts T-Ag. Expression of vascular endothelial growth factor (VEGF) receptor (VEGFR)-1, 2, Tie-l, 2 and von Willebrand factor (VWF) were assayed by reverse transcriptase-mediated polymerase chain reaction (RT-PCR). Results. We have established three cell lines incorporating 1,1′-dioctadecyl-3,3,3′,3-tetramethylindo-carbocyanine perchlorate (DiI-Ac-LDL) with a spindle shape. One of these, clone 2, strongly expressed VEGFR-2, and weakly expressed VEGFR-1 and VWF. In contrast, clone 8 showed strong expression of Tie-l, 2, and VWF, and weak expression of VEGFR-1,2. All markers were expressed strongly in clone 3. Conclusions. These data confirm that the above three TR-BME cells are novel ECs derived from bone marrow progenitors.

Original languageEnglish
Pages (from-to)9-15
Number of pages7
JournalPharmaceutical research
Volume18
Issue number1
DOIs
Publication statusPublished - 2001 Apr 25

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Keywords

  • Bone marrow
  • Cell line
  • Endothelial cell
  • Endothelial progenitor cell
  • Rat
  • SV40 large T antigen

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Medicine
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry
  • Pharmacology (medical)

Cite this

Hattori, K., Muta, M., Toi, M., Iizasa, H., Shinsei, M., Terasaki, T., Obinata, M., Ueda, M., & Nakashima, E. (2001). Establishment of bone marrow-derived endothelial cell lines from ts-SV40 T-antigen gene transgenic rats. Pharmaceutical research, 18(1), 9-15. https://doi.org/10.1023/A:1011062307374