Establishment of conditionally immortalized rat retinal pericyte cell lines (TR-rPCT) and their application in a Co-culture system using retinal capillary endothelial cell line (TR-iBRB2)1

Tetsu Kondo, Ken Ichi Hosoya, Satoko Hori, Masatoshi Tomi, Sumio Ohtsuki, Hitomi Takanaga, Emi Nakashima, Hisashi Iizasa, Tomoko Asashima, Masatsugu Ueda, Masuo Obinata, Tetsuya Terasaki

Research output: Contribution to journalArticle

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Abstract

The purpose of this study was to establish and characterize a retinal pericyte cell line from retinal capillaries of transgenic rats harboring the temperature-sensitive simian virus 40 large T-antigen gene (tsA58 Tg rat), and to apply this to the co-culture with a retinal capillary endothelial cell line. The conditionally immortalized rat retinal pericyte cell lines (TR-rPCTs), which express a temperature-sensitive large T-antigen, were obtained from two tsA58 Tg rats. These cell lines had a multicellular nodule morphology and reacted positively with von Kossa staining, a marker of calcification. TR-rPCTs cells expressed mRNA of pericyte markers such as rat intercellular adhesion molecule-1, platelet-derived growth factor-receptor β, angiopoietin-1, and osteopontin. Western blot analysis indicated that α-smooth muscle actin (α-SMA) was expressed in TR-rPCT3 and 4 cells. In contrast, α-SMA was induced by transforming growth factor-β1 and its enhancement was reduced by basic fibroblast growth factor in TR-rPCT1 and 2 cells. When TR-rPCT1 cells were cultured with a rat retinal endothelial cell line (TR-iBRB2) in a contact co-culture system, the number of TR-iBRB2 cells were significantly reduced in comparison with that of a single culture of TR-iBRB2 cells, suggesting that physical contact between pericytes and retinal endothelial cells is important for the growth of retinal endothelial cells. In conclusion, conditionally immortalized retinal pericyte cell lines were established from tsA58 Tg rats. These cell lines exhibited the properties of retinal pericytes and can be applied in co-culture systems with a retinal capillary endothelial cell line.

Original languageEnglish
Pages (from-to)145-153
Number of pages9
JournalCell Structure and Function
Volume28
Issue number3
Publication statusPublished - 2003 Jun
Externally publishedYes

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Pericytes
Coculture Techniques
Endothelial Cells
Cell Line
Viral Tumor Antigens
Angiopoietin-1
Transgenic Rats
Platelet-Derived Growth Factor Receptors
Osteopontin
Temperature
Simian virus 40
Transforming Growth Factors
Fibroblast Growth Factor 2
Smooth Muscle
Actins
Cultured Cells
Western Blotting
Staining and Labeling
Messenger RNA
Growth

Keywords

  • α-smooth muscle actin
  • Co-culture
  • Retinal endothelial cell growth
  • Retinal pericyte cell line
  • SV 40 large T-antigen

ASJC Scopus subject areas

  • Physiology
  • Structural Biology
  • Cell Biology

Cite this

Establishment of conditionally immortalized rat retinal pericyte cell lines (TR-rPCT) and their application in a Co-culture system using retinal capillary endothelial cell line (TR-iBRB2)1. / Kondo, Tetsu; Hosoya, Ken Ichi; Hori, Satoko; Tomi, Masatoshi; Ohtsuki, Sumio; Takanaga, Hitomi; Nakashima, Emi; Iizasa, Hisashi; Asashima, Tomoko; Ueda, Masatsugu; Obinata, Masuo; Terasaki, Tetsuya.

In: Cell Structure and Function, Vol. 28, No. 3, 06.2003, p. 145-153.

Research output: Contribution to journalArticle

Kondo, T, Hosoya, KI, Hori, S, Tomi, M, Ohtsuki, S, Takanaga, H, Nakashima, E, Iizasa, H, Asashima, T, Ueda, M, Obinata, M & Terasaki, T 2003, 'Establishment of conditionally immortalized rat retinal pericyte cell lines (TR-rPCT) and their application in a Co-culture system using retinal capillary endothelial cell line (TR-iBRB2)1', Cell Structure and Function, vol. 28, no. 3, pp. 145-153.
Kondo, Tetsu ; Hosoya, Ken Ichi ; Hori, Satoko ; Tomi, Masatoshi ; Ohtsuki, Sumio ; Takanaga, Hitomi ; Nakashima, Emi ; Iizasa, Hisashi ; Asashima, Tomoko ; Ueda, Masatsugu ; Obinata, Masuo ; Terasaki, Tetsuya. / Establishment of conditionally immortalized rat retinal pericyte cell lines (TR-rPCT) and their application in a Co-culture system using retinal capillary endothelial cell line (TR-iBRB2)1. In: Cell Structure and Function. 2003 ; Vol. 28, No. 3. pp. 145-153.
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abstract = "The purpose of this study was to establish and characterize a retinal pericyte cell line from retinal capillaries of transgenic rats harboring the temperature-sensitive simian virus 40 large T-antigen gene (tsA58 Tg rat), and to apply this to the co-culture with a retinal capillary endothelial cell line. The conditionally immortalized rat retinal pericyte cell lines (TR-rPCTs), which express a temperature-sensitive large T-antigen, were obtained from two tsA58 Tg rats. These cell lines had a multicellular nodule morphology and reacted positively with von Kossa staining, a marker of calcification. TR-rPCTs cells expressed mRNA of pericyte markers such as rat intercellular adhesion molecule-1, platelet-derived growth factor-receptor β, angiopoietin-1, and osteopontin. Western blot analysis indicated that α-smooth muscle actin (α-SMA) was expressed in TR-rPCT3 and 4 cells. In contrast, α-SMA was induced by transforming growth factor-β1 and its enhancement was reduced by basic fibroblast growth factor in TR-rPCT1 and 2 cells. When TR-rPCT1 cells were cultured with a rat retinal endothelial cell line (TR-iBRB2) in a contact co-culture system, the number of TR-iBRB2 cells were significantly reduced in comparison with that of a single culture of TR-iBRB2 cells, suggesting that physical contact between pericytes and retinal endothelial cells is important for the growth of retinal endothelial cells. In conclusion, conditionally immortalized retinal pericyte cell lines were established from tsA58 Tg rats. These cell lines exhibited the properties of retinal pericytes and can be applied in co-culture systems with a retinal capillary endothelial cell line.",
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AU - Hosoya, Ken Ichi

AU - Hori, Satoko

AU - Tomi, Masatoshi

AU - Ohtsuki, Sumio

AU - Takanaga, Hitomi

AU - Nakashima, Emi

AU - Iizasa, Hisashi

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AU - Ueda, Masatsugu

AU - Obinata, Masuo

AU - Terasaki, Tetsuya

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