Vol 43, No 2 (2005)
Original paper
Submitted: 2011-12-19
Published online: 2011-08-22
Autocrine growth regulation of W12 and GCA cells in culture
Anna Szuster, Magdalena Kosz-Vnenchak
Folia Histochem Cytobiol 2005;43(2):91-102.
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.
Keywords
W12 cells; GCA cells; Cell culture; Growth factors; Autocrine regulation
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
Page views
1335
Article views/downloads
864
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