Continuous maintenance of transformed fibroblasts under reduced serum conditions: Utility as a model system for investigating growth factor‐specific effects in nonquiescent cells

Kathleen M. Mulder, Michael G. Brattain

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Abstract

We report the continuous growth maintenance of untransformed and chemically transformed fibroblasts (AKR‐2B, AKR‐MCA cells) in low concentrations of serum (0.1% FBS). The cell lines established (AKR‐0.1F, MCA‐0.1F) proliferated at rates comparable to cells maintained under high serum conditions (10% FBS). Complete removal of serum from the cells did not induce quiescence. The MCA‐0.1F cells were more similar to the untransformed AKR‐2B fibroblasts in their morphology, saturation density, inability to form colonies under anchorage‐independent conditions, steady‐state level of c‐myc expression, and kinetics of induction of c‐myc in response to specific growth factors. This report demonstrates the utility of this cell line as anonquiescent model system for investigating growth factor‐specific effects in serum‐free, cycling cells. Addition of transforming growth factor‐β (TGF‐β) (5 ng/ml) to proliferating MCA‐0.IF cells, in the absence of any serum, induced a multilayered growth pattern at confluency, similar to that of AKR‐MCA cells maintained in 10% FBS. Other growth factors tested did not elicit this effect. The induction of this growth pattern by TGF‐β was associated with a sustained induction of the c‐myc proto‐oncogene at con‐fluency, but not with a restoration of anchorage‐independent growth. The data suggest that TGF‐β may play a role in the up‐regulation of c‐myc at confluency previously described for AKR‐MCA cells maintained in 10% serum.

Original languageEnglish (US)
Pages (from-to)450-458
Number of pages9
JournalJournal of Cellular Physiology
Volume138
Issue number3
DOIs
StatePublished - Mar 1989

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Fibroblasts
Maintenance
Transforming Growth Factors
Growth
Serum
Intercellular Signaling Peptides and Proteins
Cells
Restoration
Cell Line
Kinetics
Up-Regulation

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

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title = "Continuous maintenance of transformed fibroblasts under reduced serum conditions: Utility as a model system for investigating growth factor‐specific effects in nonquiescent cells",
abstract = "We report the continuous growth maintenance of untransformed and chemically transformed fibroblasts (AKR‐2B, AKR‐MCA cells) in low concentrations of serum (0.1{\%} FBS). The cell lines established (AKR‐0.1F, MCA‐0.1F) proliferated at rates comparable to cells maintained under high serum conditions (10{\%} FBS). Complete removal of serum from the cells did not induce quiescence. The MCA‐0.1F cells were more similar to the untransformed AKR‐2B fibroblasts in their morphology, saturation density, inability to form colonies under anchorage‐independent conditions, steady‐state level of c‐myc expression, and kinetics of induction of c‐myc in response to specific growth factors. This report demonstrates the utility of this cell line as anonquiescent model system for investigating growth factor‐specific effects in serum‐free, cycling cells. Addition of transforming growth factor‐β (TGF‐β) (5 ng/ml) to proliferating MCA‐0.IF cells, in the absence of any serum, induced a multilayered growth pattern at confluency, similar to that of AKR‐MCA cells maintained in 10{\%} FBS. Other growth factors tested did not elicit this effect. The induction of this growth pattern by TGF‐β was associated with a sustained induction of the c‐myc proto‐oncogene at con‐fluency, but not with a restoration of anchorage‐independent growth. The data suggest that TGF‐β may play a role in the up‐regulation of c‐myc at confluency previously described for AKR‐MCA cells maintained in 10{\%} serum.",
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T2 - Utility as a model system for investigating growth factor‐specific effects in nonquiescent cells

AU - Mulder, Kathleen M.

AU - Brattain, Michael G.

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N2 - We report the continuous growth maintenance of untransformed and chemically transformed fibroblasts (AKR‐2B, AKR‐MCA cells) in low concentrations of serum (0.1% FBS). The cell lines established (AKR‐0.1F, MCA‐0.1F) proliferated at rates comparable to cells maintained under high serum conditions (10% FBS). Complete removal of serum from the cells did not induce quiescence. The MCA‐0.1F cells were more similar to the untransformed AKR‐2B fibroblasts in their morphology, saturation density, inability to form colonies under anchorage‐independent conditions, steady‐state level of c‐myc expression, and kinetics of induction of c‐myc in response to specific growth factors. This report demonstrates the utility of this cell line as anonquiescent model system for investigating growth factor‐specific effects in serum‐free, cycling cells. Addition of transforming growth factor‐β (TGF‐β) (5 ng/ml) to proliferating MCA‐0.IF cells, in the absence of any serum, induced a multilayered growth pattern at confluency, similar to that of AKR‐MCA cells maintained in 10% FBS. Other growth factors tested did not elicit this effect. The induction of this growth pattern by TGF‐β was associated with a sustained induction of the c‐myc proto‐oncogene at con‐fluency, but not with a restoration of anchorage‐independent growth. The data suggest that TGF‐β may play a role in the up‐regulation of c‐myc at confluency previously described for AKR‐MCA cells maintained in 10% serum.

AB - We report the continuous growth maintenance of untransformed and chemically transformed fibroblasts (AKR‐2B, AKR‐MCA cells) in low concentrations of serum (0.1% FBS). The cell lines established (AKR‐0.1F, MCA‐0.1F) proliferated at rates comparable to cells maintained under high serum conditions (10% FBS). Complete removal of serum from the cells did not induce quiescence. The MCA‐0.1F cells were more similar to the untransformed AKR‐2B fibroblasts in their morphology, saturation density, inability to form colonies under anchorage‐independent conditions, steady‐state level of c‐myc expression, and kinetics of induction of c‐myc in response to specific growth factors. This report demonstrates the utility of this cell line as anonquiescent model system for investigating growth factor‐specific effects in serum‐free, cycling cells. Addition of transforming growth factor‐β (TGF‐β) (5 ng/ml) to proliferating MCA‐0.IF cells, in the absence of any serum, induced a multilayered growth pattern at confluency, similar to that of AKR‐MCA cells maintained in 10% FBS. Other growth factors tested did not elicit this effect. The induction of this growth pattern by TGF‐β was associated with a sustained induction of the c‐myc proto‐oncogene at con‐fluency, but not with a restoration of anchorage‐independent growth. The data suggest that TGF‐β may play a role in the up‐regulation of c‐myc at confluency previously described for AKR‐MCA cells maintained in 10% serum.

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