open access

Vol 43, No 2 (2005)
Original paper
Submitted: 2011-12-19
Published online: 2011-08-22
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Autocrine growth regulation of W12 and GCA cells in culture

Anna Szuster, Magdalena Kosz-Vnenchak
Folia Histochem Cytobiol 2005;43(2):91-102.

open access

Vol 43, No 2 (2005)
ORIGINAL PAPERS
Submitted: 2011-12-19
Published online: 2011-08-22

Abstract

Two rat kidney cell lines transformed by two strains of ASV virus were investigated. It was demonstrated that these two lines (1) showed density-independent growth, (2) had a decreased requirement for serum in the culture medium, (3) had the ability to grow in a chemically defined medium (without serum), and the rate of this growth had increased with the increase in starting density of cells, and (4) had the ability of anchrage-independent growth, even without serum. These results confirmed autostimulation of growth of W12 and GCA cells. It was also shown that the crude conditioned media contained autocrine growth factors, which could be extracted with 1M acetic acid. The extracts (AEs) stimulated the growth of the parental cells and NRK-49F cells almost as well as 5% calf serum and the extraction resulted in several-fold purification of mitogenic substances. These substances were not only specific to parental lines, but also stimulated growth of other transformed lines and normal NRK-49F cells. Extracts from the conditioned media of W12 and GCA cells intensified the rate of anchorage-independent growth in the concentration-dependent manner. In AE-W12, two peaks of mitogenic activity were detected (F1, F2) and similarly in AE-GCA (F3, F4). Fractions F2 (~ 8 kDa), F3 (~25 kDa) and F4 (~ 12 kDa) were thermostable but F1 (~ 45 kDa) was thermolabile. All four fractions were sensitive to trypsin and DTT treatment, and were acid-stable. Using ELISA kit it was shown that W12 and GCA cells released TGFβ1 and GCA cells released very small quantities of bFGF. These results confirmed the autocrine regulation of growth in both cell lines.

Abstract

Two rat kidney cell lines transformed by two strains of ASV virus were investigated. It was demonstrated that these two lines (1) showed density-independent growth, (2) had a decreased requirement for serum in the culture medium, (3) had the ability to grow in a chemically defined medium (without serum), and the rate of this growth had increased with the increase in starting density of cells, and (4) had the ability of anchrage-independent growth, even without serum. These results confirmed autostimulation of growth of W12 and GCA cells. It was also shown that the crude conditioned media contained autocrine growth factors, which could be extracted with 1M acetic acid. The extracts (AEs) stimulated the growth of the parental cells and NRK-49F cells almost as well as 5% calf serum and the extraction resulted in several-fold purification of mitogenic substances. These substances were not only specific to parental lines, but also stimulated growth of other transformed lines and normal NRK-49F cells. Extracts from the conditioned media of W12 and GCA cells intensified the rate of anchorage-independent growth in the concentration-dependent manner. In AE-W12, two peaks of mitogenic activity were detected (F1, F2) and similarly in AE-GCA (F3, F4). Fractions F2 (~ 8 kDa), F3 (~25 kDa) and F4 (~ 12 kDa) were thermostable but F1 (~ 45 kDa) was thermolabile. All four fractions were sensitive to trypsin and DTT treatment, and were acid-stable. Using ELISA kit it was shown that W12 and GCA cells released TGFβ1 and GCA cells released very small quantities of bFGF. These results confirmed the autocrine regulation of growth in both cell lines.
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Keywords

W12 cells; GCA cells; Cell culture; Growth factors; Autocrine regulation

About this article
Title

Autocrine growth regulation of W12 and GCA cells in culture

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 43, No 2 (2005)

Article type

Original paper

Pages

91-102

Published online

2011-08-22

Bibliographic record

Folia Histochem Cytobiol 2005;43(2):91-102.

Keywords

W12 cells
GCA cells
Cell culture
Growth factors
Autocrine regulation

Authors

Anna Szuster
Magdalena Kosz-Vnenchak

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