TGF-α is a potent mitogen for primary cultures of guinea pig gastric mucous epithelial cells

Michael Rutten, P. J. Dempsey, T. E. Solomon, R. J. Coffey

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

Transforming growth factor-α (TGF-α) and epidermal growth factor (EGF) are thought to be important in gastric epithelial proliferation and repair. It was therefore of interest to determine if TGF-α and EGF promoted the growth of an in vitro primary culture system of guinea pig gastric mucous epithelial cells (MEC). MEC were isolated from guinea pig stomachs and cultured in 24-well Primaria plates with DMEM with or without 10% fetal calf serum (FCS). Growth of MEC was determined by changes in [3H]thymidine uptake, cell counts, protein, and DNA. The sources of peptides were human recombinant TGF-α (recTGF-α) and human recombinant EGF (recEGF). Both recTGF-α and recEGF were used at equipotent doses as determined by competing activity in a 125I-labeled TGF-α radioreceptor binding assay using A-431 cells. Basal growth (no peptides) of MEC in 10% FCS was dependent on the initial plating density. Under serum-free conditions, [3H]thymidine uptake increased up to 17-fold at 24 h with recTGF-α (0.1-10.0 nM) compared with only a 4-fold increase using rec-EGF (0.1-10.0 nM) at this same time period. Under serum-free conditions, recTGF-α (0.01-10.0 nM) increased cell counts up to 4.9-fold over control cultures, whereas similar does of recEGF produced a 2.5-fold increase in cell counts. Administration of recEGF (1 ng/ml) resulted in a 1.9-fold increase in the 4.8-kb TGF-α mRNA transcript, and TGF-α protein immunoreactivity was found in both 24-h conditioned media and cell lysates. We conclude that TGF-α is a more potent mitogen for guinea pig gastric mucous cells, and the data suggest that TGF-α may act as an autocrine factor for these cells.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume265
Issue number2 28-2
StatePublished - 1993

Fingerprint

Transforming Growth Factors
Mitogens
Epidermal Growth Factor
Stomach
Guinea Pigs
Epithelial Cells
Cell Count
Serum
Thymidine
Growth
Radioligand Assay
Peptides
Conditioned Culture Medium
Proteins
Messenger RNA
DNA

Keywords

  • epidermal growth factor
  • growth
  • repair
  • stomach
  • ulcers

ASJC Scopus subject areas

  • Physiology
  • Gastroenterology

Cite this

TGF-α is a potent mitogen for primary cultures of guinea pig gastric mucous epithelial cells. / Rutten, Michael; Dempsey, P. J.; Solomon, T. E.; Coffey, R. J.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 265, No. 2 28-2, 1993.

Research output: Contribution to journalArticle

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AB - Transforming growth factor-α (TGF-α) and epidermal growth factor (EGF) are thought to be important in gastric epithelial proliferation and repair. It was therefore of interest to determine if TGF-α and EGF promoted the growth of an in vitro primary culture system of guinea pig gastric mucous epithelial cells (MEC). MEC were isolated from guinea pig stomachs and cultured in 24-well Primaria plates with DMEM with or without 10% fetal calf serum (FCS). Growth of MEC was determined by changes in [3H]thymidine uptake, cell counts, protein, and DNA. The sources of peptides were human recombinant TGF-α (recTGF-α) and human recombinant EGF (recEGF). Both recTGF-α and recEGF were used at equipotent doses as determined by competing activity in a 125I-labeled TGF-α radioreceptor binding assay using A-431 cells. Basal growth (no peptides) of MEC in 10% FCS was dependent on the initial plating density. Under serum-free conditions, [3H]thymidine uptake increased up to 17-fold at 24 h with recTGF-α (0.1-10.0 nM) compared with only a 4-fold increase using rec-EGF (0.1-10.0 nM) at this same time period. Under serum-free conditions, recTGF-α (0.01-10.0 nM) increased cell counts up to 4.9-fold over control cultures, whereas similar does of recEGF produced a 2.5-fold increase in cell counts. Administration of recEGF (1 ng/ml) resulted in a 1.9-fold increase in the 4.8-kb TGF-α mRNA transcript, and TGF-α protein immunoreactivity was found in both 24-h conditioned media and cell lysates. We conclude that TGF-α is a more potent mitogen for guinea pig gastric mucous cells, and the data suggest that TGF-α may act as an autocrine factor for these cells.

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