Development of a bioassay for pulmonary cell production of fibrogenic factors

R. Reist, K. Bryner, P. Wearden, J. Blackford, Kent Vrana, V. Castranova, R. Dey

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Summary: Fibroblast proliferation and enhancement of collagen synthesis are key steps in the development and progression of pulmonary fibrosis. The current investigation presents a fibroblast proliferation assay to detect the release of competence factors from pneumocytes. The specific objective of this study was to define cell culture conditions for fibroblasts that would allow selective detection of competence factors in test media. The results indicate the following: (a) fibroblasts should be cultured for 2 days in plasma-free medium rather than 2% plasma to assure complete quiescence; (b) 1% plasma was optimum for producing progression activity for the bioassay of a competence factor, while 0.5% plasma did not produce sufficient progression activity (yielding false negatives) and 2% plasma often contained significant competence activity (yielding false positives); and (c) fresh plasma should be used since the progression activity of plasma was adversely affected by freezing and thawing. Under these conditions, this bioassay can be employed with either rat or human lung fibroblasts to detect competence factors, such as platelet-derived growth factor (PDGF), at levels as low as 25 ng/ml.

Original languageEnglish (US)
Pages (from-to)53-65
Number of pages13
JournalToxicology Mechanisms and Methods
Volume1
Issue number1
DOIs
StatePublished - Jan 1 1991

Fingerprint

Bioassay
Biological Assay
Fibroblasts
Mental Competency
Plasmas
Lung
Thermodynamic properties
Alveolar Epithelial Cells
Thawing
Pulmonary Fibrosis
Platelet-Derived Growth Factor
Cell culture
Freezing
Rats
Assays
Collagen
Cell Culture Techniques

All Science Journal Classification (ASJC) codes

  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Reist, R. ; Bryner, K. ; Wearden, P. ; Blackford, J. ; Vrana, Kent ; Castranova, V. ; Dey, R. / Development of a bioassay for pulmonary cell production of fibrogenic factors. In: Toxicology Mechanisms and Methods. 1991 ; Vol. 1, No. 1. pp. 53-65.
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Reist, R, Bryner, K, Wearden, P, Blackford, J, Vrana, K, Castranova, V & Dey, R 1991, 'Development of a bioassay for pulmonary cell production of fibrogenic factors', Toxicology Mechanisms and Methods, vol. 1, no. 1, pp. 53-65. https://doi.org/10.3109/15376519109036525

Development of a bioassay for pulmonary cell production of fibrogenic factors. / Reist, R.; Bryner, K.; Wearden, P.; Blackford, J.; Vrana, Kent; Castranova, V.; Dey, R.

In: Toxicology Mechanisms and Methods, Vol. 1, No. 1, 01.01.1991, p. 53-65.

Research output: Contribution to journalArticle

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AU - Reist, R.

AU - Bryner, K.

AU - Wearden, P.

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AU - Castranova, V.

AU - Dey, R.

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N2 - Summary: Fibroblast proliferation and enhancement of collagen synthesis are key steps in the development and progression of pulmonary fibrosis. The current investigation presents a fibroblast proliferation assay to detect the release of competence factors from pneumocytes. The specific objective of this study was to define cell culture conditions for fibroblasts that would allow selective detection of competence factors in test media. The results indicate the following: (a) fibroblasts should be cultured for 2 days in plasma-free medium rather than 2% plasma to assure complete quiescence; (b) 1% plasma was optimum for producing progression activity for the bioassay of a competence factor, while 0.5% plasma did not produce sufficient progression activity (yielding false negatives) and 2% plasma often contained significant competence activity (yielding false positives); and (c) fresh plasma should be used since the progression activity of plasma was adversely affected by freezing and thawing. Under these conditions, this bioassay can be employed with either rat or human lung fibroblasts to detect competence factors, such as platelet-derived growth factor (PDGF), at levels as low as 25 ng/ml.

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