Secretion of insulin-like growth factor binding protein-1 from individual hepatocytes

G. Eghbali-Fatourechi, Cheryl A Conover, Gary C Sieck, Gregory James Gores, L. A. Fitzpatrick

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The reverse hemolytic plaque assay (RHPA) uses complement-mediated red blood cell lysis to detect peptide secretion by individual cells. Initially, the RHPA was used to study the function of neurons and B lymphocytes. More recently, the RHPA has been adapted to measure hormone release from individual pituitary, parathyroid, luteal, and pancreatic islet cells. We have applied this technique to detect insulin-like growth factor binding protein-1 (IGFBP-1) secretion by a human hepatoma cell line (HepG2). We proposed that the technique of RHPA could be used to study peptide release from single hepatocytes in various defined conditions. Our goal was the study of the kinetics of IGFBP-1 secretion from hepatoma cells and rat hepatocytes and to determine the heterogeneity of the cell population regarding the secretion of IGFBP-1. To evaluate the optimal conditions of IGFBP-1 secretion by hepatoma cells and rat hepatocytes and to evaluate the influence of cell dispersion on hepatocyte's behavior, we evaluated three techniques of cell dispersion: trypsin digestion, collagenase digestion, and mechanical dispersion. We tested cell viability, determined the percentage of secreting cells versus non-secreting cells, and measured mean plaque area which is a function of the amount of IGFBP-1 secreted by an individual cell. We determined the optimal IGFBP-1 antibody dilution for the detection of secreted IGFBP-1 by hepatocytes, evaluated the initiation of IGFBP-1 secretion from cultured cells, and quantified time-dependent IGFBP-1 secretion. In addition to demonstrating the feasibility of measuring IGFBP-1 from a cultured cell line, we measured IGFBP-1 release from freshly dispersed rat hepatocytes.

Original languageEnglish (US)
Pages (from-to)243-259
Number of pages17
JournalResearch Communications in Molecular Pathology and Pharmacology
Volume85
Issue number3
StatePublished - 1994

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Insulin-Like Growth Factor Binding Protein 1
Hepatocytes
Cells
Assays
Rats
Hepatocellular Carcinoma
Islets of Langerhans
Hemolytic Plaque Technique
Digestion
Cultured Cells
Cell Line
Peptides
Lymphocytes
Corpus Luteum
Collagenases
Population Characteristics
Trypsin
Dilution
Neurons
Cell Survival

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Secretion of insulin-like growth factor binding protein-1 from individual hepatocytes. / Eghbali-Fatourechi, G.; Conover, Cheryl A; Sieck, Gary C; Gores, Gregory James; Fitzpatrick, L. A.

In: Research Communications in Molecular Pathology and Pharmacology, Vol. 85, No. 3, 1994, p. 243-259.

Research output: Contribution to journalArticle

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