Selective migration of α-smooth muscle actin-positive myofibroblasts toward fibronectin in the Boyden's blindwell chamber

Masashi Kawamoto, Takakuni Matsunami, Ronald F. Ertl, Yuh Fukuda, Maki Ogawa, John R. Spurzem, Nobuaki Yamanaka, Stephen I. Rennard

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

1. In order to address the hypothesis that migrating fibroblasts have a different phenotype, human fetal lung fibroblasts (HFL-1) cells were evaluated in the Boyden blindwell chamber migration assay followed by immunoelectron microscopy. 2. HFL-1 cells were placed on nucleopore filters and incubated for 2 h using purified human plasma fibronectin (pFn) as a chemoattractant. Filters were then processed for immunoelectron microscopy using antibodies for α-smooth muscle (α-SM) actin as a marker for myofibroblasts, cellular fibronectin (cFn) and VLA-5. 3. Cells which had migrated to the bottom side of the filter were more likely to express α-SM actin, 29.1 ± 3.4% of cells, compared with cells which did not migrate through the filter, 12.4 ± 1.3% (P < 0.05). The total proportion of α-SM-actin-positive cells located on both sides of the filter showed no difference between those which had migrated toward pFn and controls (17.7 ± 1.0% compared with 20.2 ± 2.5%). 4. cFn-positive cells showed minimal differences compared with control cells, while perinuclear and endoplasmic reticulum staining of VLA-5 was observed only in the cells treated with pFn. 5. The results show that HFL-1 cells are heterogenous for α-SM actin expression. Short-term incubation, with pFn did not change the proportion of α-SM-actin-positive HFL-1 cells. Cells which migrate, however, are enriched for α-SM actin expression, pFn-induced fibroblast chemotaxis can selectively recruit myofibroblasts with increased α-SM actin expression, a feature which may contribute to the altered population of cells at sites of fibrosis.

Original languageEnglish (US)
Pages (from-to)355-362
Number of pages8
JournalClinical Science
Volume93
Issue number4
DOIs
StatePublished - Jan 1 1997

Fingerprint

Myofibroblasts
Fibronectins
Smooth Muscle
Actins
Integrin alpha5beta1
Immunoelectron Microscopy
Fibroblasts
Chemotactic Factors
Chemotaxis
Endoplasmic Reticulum
Fibrosis

Keywords

  • Boyden chamber
  • Chemotaxis
  • Fibronectin
  • Myofibroblast
  • α-smooth muscle actin

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Kawamoto, M., Matsunami, T., Ertl, R. F., Fukuda, Y., Ogawa, M., Spurzem, J. R., ... Rennard, S. I. (1997). Selective migration of α-smooth muscle actin-positive myofibroblasts toward fibronectin in the Boyden's blindwell chamber. Clinical Science, 93(4), 355-362. https://doi.org/10.1042/cs0930355

Selective migration of α-smooth muscle actin-positive myofibroblasts toward fibronectin in the Boyden's blindwell chamber. / Kawamoto, Masashi; Matsunami, Takakuni; Ertl, Ronald F.; Fukuda, Yuh; Ogawa, Maki; Spurzem, John R.; Yamanaka, Nobuaki; Rennard, Stephen I.

In: Clinical Science, Vol. 93, No. 4, 01.01.1997, p. 355-362.

Research output: Contribution to journalArticle

Kawamoto, M, Matsunami, T, Ertl, RF, Fukuda, Y, Ogawa, M, Spurzem, JR, Yamanaka, N & Rennard, SI 1997, 'Selective migration of α-smooth muscle actin-positive myofibroblasts toward fibronectin in the Boyden's blindwell chamber', Clinical Science, vol. 93, no. 4, pp. 355-362. https://doi.org/10.1042/cs0930355
Kawamoto, Masashi ; Matsunami, Takakuni ; Ertl, Ronald F. ; Fukuda, Yuh ; Ogawa, Maki ; Spurzem, John R. ; Yamanaka, Nobuaki ; Rennard, Stephen I. / Selective migration of α-smooth muscle actin-positive myofibroblasts toward fibronectin in the Boyden's blindwell chamber. In: Clinical Science. 1997 ; Vol. 93, No. 4. pp. 355-362.
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abstract = "1. In order to address the hypothesis that migrating fibroblasts have a different phenotype, human fetal lung fibroblasts (HFL-1) cells were evaluated in the Boyden blindwell chamber migration assay followed by immunoelectron microscopy. 2. HFL-1 cells were placed on nucleopore filters and incubated for 2 h using purified human plasma fibronectin (pFn) as a chemoattractant. Filters were then processed for immunoelectron microscopy using antibodies for α-smooth muscle (α-SM) actin as a marker for myofibroblasts, cellular fibronectin (cFn) and VLA-5. 3. Cells which had migrated to the bottom side of the filter were more likely to express α-SM actin, 29.1 ± 3.4{\%} of cells, compared with cells which did not migrate through the filter, 12.4 ± 1.3{\%} (P < 0.05). The total proportion of α-SM-actin-positive cells located on both sides of the filter showed no difference between those which had migrated toward pFn and controls (17.7 ± 1.0{\%} compared with 20.2 ± 2.5{\%}). 4. cFn-positive cells showed minimal differences compared with control cells, while perinuclear and endoplasmic reticulum staining of VLA-5 was observed only in the cells treated with pFn. 5. The results show that HFL-1 cells are heterogenous for α-SM actin expression. Short-term incubation, with pFn did not change the proportion of α-SM-actin-positive HFL-1 cells. Cells which migrate, however, are enriched for α-SM actin expression, pFn-induced fibroblast chemotaxis can selectively recruit myofibroblasts with increased α-SM actin expression, a feature which may contribute to the altered population of cells at sites of fibrosis.",
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AU - Rennard, Stephen I.

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AB - 1. In order to address the hypothesis that migrating fibroblasts have a different phenotype, human fetal lung fibroblasts (HFL-1) cells were evaluated in the Boyden blindwell chamber migration assay followed by immunoelectron microscopy. 2. HFL-1 cells were placed on nucleopore filters and incubated for 2 h using purified human plasma fibronectin (pFn) as a chemoattractant. Filters were then processed for immunoelectron microscopy using antibodies for α-smooth muscle (α-SM) actin as a marker for myofibroblasts, cellular fibronectin (cFn) and VLA-5. 3. Cells which had migrated to the bottom side of the filter were more likely to express α-SM actin, 29.1 ± 3.4% of cells, compared with cells which did not migrate through the filter, 12.4 ± 1.3% (P < 0.05). The total proportion of α-SM-actin-positive cells located on both sides of the filter showed no difference between those which had migrated toward pFn and controls (17.7 ± 1.0% compared with 20.2 ± 2.5%). 4. cFn-positive cells showed minimal differences compared with control cells, while perinuclear and endoplasmic reticulum staining of VLA-5 was observed only in the cells treated with pFn. 5. The results show that HFL-1 cells are heterogenous for α-SM actin expression. Short-term incubation, with pFn did not change the proportion of α-SM-actin-positive HFL-1 cells. Cells which migrate, however, are enriched for α-SM actin expression, pFn-induced fibroblast chemotaxis can selectively recruit myofibroblasts with increased α-SM actin expression, a feature which may contribute to the altered population of cells at sites of fibrosis.

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