Characterization of SV40-transfected cell strains from rabbit keratocytes

Patricia A. Barry-Lane, Steven E. Wilson, H. Dwight Cavanagh, W. Matthew Petroll, James V. Jester

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The process of corneal wound healing involves the transformation of adjacent corneal keratocytes to myofibroblast-like cells characterized by the development of prominent microfilament bundles containing α-smooth muscle- specific actin (α-SM), a contractile protein thought to be important in mediating wound contraction. Recent studies have shown that the expression of α-SM in cultured corneal keratocytes can be induced by serum and TGF(β1). To study the cellular and molecular mechanisms underlying this transformation process and to begin to identify the role of α-SM in wound contractile events, we generated immortalized rabbit corneal cell strains with extended life by using SV40 transfection. Two unique strains were isolated (TRK-36 and TRK-43). TRK-36, which appears similar to normal corneal keratocytes, maintains a stellate, keratocyte morphology when grown in the absence of serum and transforms to a myofibroblast-like cell when treated with TGF(β1) (1 ng/ml), as indicated by the induced expression of α-SM actin. TRK-43 exhibits features characteristic of myofibroblasts in that it constitutively expresses α-SM actin under serum-free conditions. Both strains show in vitro contraction of collagen gels ≤80% in 24 h in serum-containing medium. Interestingly, under serum-free conditions, TRK-43 cells showed significantly greater contraction of collagen gels compared with those of TRK-36. Overall, the establishment and further study of these cell strains may provide important insights into the molecular mechanisms underlying myofibroblast transformation.

Original languageEnglish (US)
Pages (from-to)72-78
Number of pages7
JournalCornea
Volume16
Issue number1
StatePublished - Jan 1997

Fingerprint

Corneal Keratocytes
Myofibroblasts
Rabbits
Actins
Serum
Collagen
Gels
Contractile Proteins
Wounds and Injuries
Actin Cytoskeleton
Wound Healing
Transfection
Smooth Muscle

Keywords

  • α-Smooth muscle
  • Corneal keratocytes
  • Myofibroblast
  • specific actin
  • SV40

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Barry-Lane, P. A., Wilson, S. E., Cavanagh, H. D., Petroll, W. M., & Jester, J. V. (1997). Characterization of SV40-transfected cell strains from rabbit keratocytes. Cornea, 16(1), 72-78.

Characterization of SV40-transfected cell strains from rabbit keratocytes. / Barry-Lane, Patricia A.; Wilson, Steven E.; Cavanagh, H. Dwight; Petroll, W. Matthew; Jester, James V.

In: Cornea, Vol. 16, No. 1, 01.1997, p. 72-78.

Research output: Contribution to journalArticle

Barry-Lane, PA, Wilson, SE, Cavanagh, HD, Petroll, WM & Jester, JV 1997, 'Characterization of SV40-transfected cell strains from rabbit keratocytes', Cornea, vol. 16, no. 1, pp. 72-78.
Barry-Lane, Patricia A. ; Wilson, Steven E. ; Cavanagh, H. Dwight ; Petroll, W. Matthew ; Jester, James V. / Characterization of SV40-transfected cell strains from rabbit keratocytes. In: Cornea. 1997 ; Vol. 16, No. 1. pp. 72-78.
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