Proliferation of a human embryonal carcinoma-derived cell line in serum-free medium: inter-relationship between growth factor requirements and membrane receptor expression

1985 ◽  
Vol 73 (1) ◽  
pp. 361-373
Author(s):  
W. Engstrom ◽  
A.R. Rees ◽  
J.K. Heath
Keyword(s):  
Growth Factor ◽  
Cell Line ◽  
Embryonal Carcinoma ◽  
Density Lipoprotein ◽  
Basal Medium ◽  
Receptor Expression ◽  
Embryonal Carcinoma Cell ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free

Substantial multiplication in vitro of cloned cells from a human embryonal carcinoma cell line, Tera 2, has been obtained in a basal medium (DMEM/Ham's F12,50:50, v/v) supplemented with 10 micrograms low density lipoprotein/ml, 100 micrograms high density lipoprotein/ml, 100 ng multiplication stimulating activity/ml, 100 ng insulin/ml and 1 microgram transferrin/ml. The growth rate appears to be similar to that obtained in 10% serum. Furthermore, studies on the expression of cell surface receptors revealed that cloned Tera 2 cells express high-affinity receptors for IGF-II but not for insulin. The cells also express receptors for Epidermal Growth Factor (EGF) even though the addition of EGF does not stimulate their proliferation in serum-free medium. These results suggest that the expression of specific growth factor receptors is not an absolute determinant of hormone responsiveness.

scholarly journals Nerve growth factor prevents the death and stimulates the neuronal differentiation of clonal PC12 pheochromocytoma cells in serum-free medium

1978 ◽  
Vol 78 (3) ◽  
pp. 747-755 ◽  
Author(s):  
LA Greene
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Neurite Outgrowth ◽  
Cell Line ◽  
Pc12 Cells ◽  
Serum Free Medium ◽  
Free Medium ◽  
Nerve Growth ◽  
Serum Free ◽  
Serum Free Conditions

The PC12 clone is a noradrenergic cell line derived from a rat pheochromocytoma. In culture medium containing horse serum, PC12 cells undergo mitosis; when nerve growth factor (NGF) is included in the medium, the cells cease multiplication and extend neuritis. It is shown here: (a) that PC12 cells are not viable in serum-free medium. When serum is withdrawn, 90 percent of the cells die within 4-6 days and 99 percent by 2-3 wk. (b) If NGF is added at the time of serum withdrawal, the cells undergo one doubling and remain viable for at least 1 mo. (c) Addition of NGF to cultures after more than 2 days in serum-free conditions results in maintenance of surviving cells, but not in an increase in cell number. (d) NGD also induces neurite outgrowth from PC12 cells in serum-free medium. (e) NGF-treated cells exhibit much less cell-cell and neurite-neurite aggregation in the absence than in the presence of serum. (f) The apparent minimum level of 2.5S NGF required for PC12 survival and morphological differentiation in serum-free medium is about 10 ng/ml (approximately 0.4 nM). (g) Withdrawal of NGF in serum-free conditions results in degeneration of neurites and loss of cell viability. (h) Experiments with campotothecin demonstrate that the effects of NGF on survival and neurite outgrowth may be uncoupled and suggest that the survival effects are transcriptionally independent. The present results also suggest that PC12 cells have a requirement for NGF (similar to that of normal sympathetic neurons) and that serum may substitute for this requirement. In addition, the present system of maintaining a highly differentiated cell line in a chemically defined medium suggests certain experimental opportunities.

scholarly journals The effects of various hormones and growth factors on the growth of human insulin-producing cell line in serum-free medium

1997 ◽  
Vol 29 (4) ◽  
pp. 209-216 ◽  
Author(s):  
Jae-Jeong Lee ◽  
Jai-Hyun Kwon ◽  
Yong Keun Park ◽  
Ohoak Kwon ◽  
Tai-Wook Yoon
Keyword(s):  
Growth Factors ◽  
Cell Line ◽  
Human Insulin ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free

Fibroblast growth factor-2 in serum-free medium is a potent mitogen and reduces dedifferentiation of human ear chondrocytes in monolayer culture

2004 ◽  
Vol 23 (4) ◽  
pp. 231-241 ◽  
Author(s):  
E.W. Mandl ◽  
H. Jahr ◽  
J.L.M. Koevoet ◽  
J.P.T.M. van Leeuwen ◽  
H. Weinans ◽  
...  
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Monolayer Culture ◽  
Fibroblast Growth Factor 2 ◽  
Fibroblast Growth ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Human Ear

The K-fgf/hst oncogene induces transformation through an autocrine mechanism that requires extracellular stimulation of the mitogenic pathway

1991 ◽  
Vol 11 (2) ◽  
pp. 1138-1145
Author(s):  
D Talarico ◽  
C Basilico
Keyword(s):  
Growth Factor ◽  
Receptor Activation ◽  
Soft Agar ◽  
Serum Free Medium ◽  
Free Medium ◽  
3T3 Cells ◽  
Serum Free ◽  
Nih 3T3 ◽  
Autocrine Mechanism ◽  
Nih 3T3 Cells

The K-fgf/hst oncogene encodes a secreted growth factor of the fibroblast growth factor (FGF) family. The ability of K-fgf-transformed cells to grow in soft agar and in serum-free medium is inhibited by anti-K-FGF neutralizing antibodies, consistent with an autocrine mechanism of transformation. The transformed properties of clones that express high levels of K-FGF are, however, only partially affected. To better define the autocrine mechanism of transformation by K-fgf and to determine whether receptor activation could occur intracellularly, we constructed two mutants of the K-fgf cDNA. Deletion of the sequences encoding the signal peptide suppressed K-fgf ability to induce foci in NIH 3T3 cells. A few morphologically transformed colonies were observed in cotransfection experiments, and they were found to express high levels of cytoplasmic K-FGF. However, their ability to grow in serum-free medium and in soft agar was inhibited by anti-K-FGF antibodies. Addition of a sequence encoding the KDEL endoplasmic reticulum and Golgi retention signal to the K-fgf cDNA led to accumulation of the growth factor in intracellular compartments. The ability of the KDEL mutant to induce foci in NIH 3T3 cells was much lower than that of the wild-type cDNA, and also in this case the transformed phenotype was reverted by anti-K-FGF antibodies. These and other findings indicate that the transformed phenotype of cells expressing a nonsecretory K-FGF is due to the extracellular activation of the receptor by the small amounts of growth factor that these cells still release. Thus, transformation by K-fgf appears to be due to an autocrine growth mechanisms that requires activation of the mitogenic pathway at the cell surface.

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