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

Research output: Contribution to journalArticle

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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

Fingerprint

Transgenic Rats
Polyomavirus Transforming Antigens
Endothelial cells
Rats
Bone
Endothelial Cells
Genes
Bone Marrow
Vascular Endothelial Growth Factor Receptor
von Willebrand Factor
Cell Line
Clone Cells
Carbocyanines
Vascular Endothelial Growth Factor Receptor-1
Vascular Endothelial Growth Factor Receptor-2
Temperature
Polymerase chain reaction
RNA-Directed DNA Polymerase
Viral Tumor Antigens
Reverse Transcriptase Polymerase Chain Reaction

Keywords

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

ASJC Scopus subject areas

  • Chemistry(all)
  • Pharmaceutical Science
  • Pharmacology

Cite this

Hattori, K., Muta, M., Toi, M., Iizasa, H., Shinsei, M., Terasaki, T., ... 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

Establishment of bone marrow-derived endothelial cell lines from ts-SV40 T-antigen gene transgenic rats. / Hattori, K.; Muta, M.; Toi, M.; Iizasa, H.; Shinsei, M.; Terasaki, T.; Obinata, M.; Ueda, M.; Nakashima, E.

In: Pharmaceutical Research, Vol. 18, No. 1, 2001, p. 9-15.

Research output: Contribution to journalArticle

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, vol. 18, no. 1, pp. 9-15. https://doi.org/10.1023/A:1011062307374
Hattori, K. ; Muta, M. ; Toi, M. ; Iizasa, H. ; Shinsei, M. ; Terasaki, T. ; Obinata, M. ; Ueda, M. ; Nakashima, E. / Establishment of bone marrow-derived endothelial cell lines from ts-SV40 T-antigen gene transgenic rats. In: Pharmaceutical Research. 2001 ; Vol. 18, No. 1. pp. 9-15.
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AU - Hattori, K.

AU - Muta, M.

AU - Toi, M.

AU - Iizasa, H.

AU - Shinsei, M.

AU - Terasaki, T.

AU - Obinata, M.

AU - Ueda, M.

AU - Nakashima, E.

PY - 2001

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N2 - 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.

AB - 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.

KW - Bone marrow

KW - Cell line

KW - Endothelial cell

KW - Endothelial progenitor cell

KW - Rat

KW - SV40 large T antigen

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