Improved clonal and nonclonal growth of human, rat and bovine adrenocortical cells in culture

Janette McAllister, Peter J. Hornsby

Research output: Contribution to journalReview article

39 Citations (Scopus)

Abstract

This report describes the development of a culture system for long-term growth and cloning of human fetal adrenocortical cells. Optimal conditions for stimulating clonal growth were determned by testing the efficacy of horse serum (HS), fetal bovine serum (FBS), fibroblast growth factor (FGF), epidermal growth factor (EGF), fibronectin, and a combination of growth factors, UltroSer G, in stimulating growth from low density. Optimal conditions for clonal growth were achieved using fibronectin-coated dishes and DME/F12 medium with 10% FEBS, 10% HS, 2% UltroSer G, and 100 ng/ml FGF or 100 p M EGF. Conditions for growth at clonal density were found to be optimal for growth of early passage, nonclonal cultures at higher densities. The improved growth conditions used for cloning were shown to allow continued long-term growth of nonclonal human adrenocortical cells without fibroblasts overgrowth. All cells in cultures grown in HS, FBS, and UltroSer G had morphologic characteristics of adrenocortical cells, whereas cells grown in FBS only rapidly became overgrown with fibroblasts. Clonal and nonclonal early passage human adrenocortical cells had smilar mitogenic responses to FGF and EGF. Whereas FGF, EGF, and UltroSer G showed similar stimulation of DNA synthesis and clonal growth in human adrenocortical cells and human adrenal gland fibroblasts, the tumor promoter 12-O-teradecanoylphorbol-13-acetate stimulated growth only in adrenocortical cells and was strongly inhibitory to growth in fibroblasts. In both cell types, forskolin inhibited DNA synthesis. Human adrenocortical cell cultures were functional and synthesized cortisol, dehydroepiandrosterone, and dehydroepiandrosterone sulfate. The improved growth conditions for clonal growth of human adrenocortial cells also provided optimal conditions for long-term growth of cultured rat adrenocortical cells and ncreased the cloning efficiency of cultured bovine adrenocortical cells.

Original languageEnglish (US)
Pages (from-to)677-685
Number of pages9
JournalIn Vitro Cellular & Developmental Biology
Volume23
Issue number10
DOIs
StatePublished - Oct 1 1987

Fingerprint

Rats
Cell Culture Techniques
Growth
Fibroblast Growth Factors
Fibroblasts
Epidermal Growth Factor
Cloning
Serum
Horses
Organism Cloning
Cells
Fibronectins
Cell culture
Dehydroepiandrosterone Sulfate
Dehydroepiandrosterone
DNA
Colforsin
Adrenal Glands
Carcinogens
Hydrocortisone

All Science Journal Classification (ASJC) codes

  • Developmental Biology
  • Cell Biology
  • Medicine(all)

Cite this

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title = "Improved clonal and nonclonal growth of human, rat and bovine adrenocortical cells in culture",
abstract = "This report describes the development of a culture system for long-term growth and cloning of human fetal adrenocortical cells. Optimal conditions for stimulating clonal growth were determned by testing the efficacy of horse serum (HS), fetal bovine serum (FBS), fibroblast growth factor (FGF), epidermal growth factor (EGF), fibronectin, and a combination of growth factors, UltroSer G, in stimulating growth from low density. Optimal conditions for clonal growth were achieved using fibronectin-coated dishes and DME/F12 medium with 10{\%} FEBS, 10{\%} HS, 2{\%} UltroSer G, and 100 ng/ml FGF or 100 p M EGF. Conditions for growth at clonal density were found to be optimal for growth of early passage, nonclonal cultures at higher densities. The improved growth conditions used for cloning were shown to allow continued long-term growth of nonclonal human adrenocortical cells without fibroblasts overgrowth. All cells in cultures grown in HS, FBS, and UltroSer G had morphologic characteristics of adrenocortical cells, whereas cells grown in FBS only rapidly became overgrown with fibroblasts. Clonal and nonclonal early passage human adrenocortical cells had smilar mitogenic responses to FGF and EGF. Whereas FGF, EGF, and UltroSer G showed similar stimulation of DNA synthesis and clonal growth in human adrenocortical cells and human adrenal gland fibroblasts, the tumor promoter 12-O-teradecanoylphorbol-13-acetate stimulated growth only in adrenocortical cells and was strongly inhibitory to growth in fibroblasts. In both cell types, forskolin inhibited DNA synthesis. Human adrenocortical cell cultures were functional and synthesized cortisol, dehydroepiandrosterone, and dehydroepiandrosterone sulfate. The improved growth conditions for clonal growth of human adrenocortial cells also provided optimal conditions for long-term growth of cultured rat adrenocortical cells and ncreased the cloning efficiency of cultured bovine adrenocortical cells.",
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Improved clonal and nonclonal growth of human, rat and bovine adrenocortical cells in culture. / McAllister, Janette; Hornsby, Peter J.

In: In Vitro Cellular & Developmental Biology, Vol. 23, No. 10, 01.10.1987, p. 677-685.

Research output: Contribution to journalReview article

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