scholarly journals One-week 96-well soft agar growth assay for cancer target validation

BioTechniques ◽  
2004 ◽  
Vol 36 (5) ◽  
pp. 826-833 ◽  
Author(s):  
Ning Ke ◽  
Aaron Albers ◽  
Gisela Claassen ◽  
De-hua Yu ◽  
Jon E. Chatterton ◽  
...  
Keyword(s):  
Soft Agar ◽  
Target Validation ◽  
Soft Agar Growth ◽  
Growth Assay

Effects of antiinflammatory drugs on soft agar growth of rabbit articular chondrocytes

1989 ◽  
Vol 18 (3) ◽  
pp. 7-10 ◽  
Author(s):  
Yukio Kato ◽  
Katsuhiko Sato ◽  
Tatsuya Koike ◽  
Kazuhisha Nakashima ◽  
Wei Qun Yen ◽  
...  
Keyword(s):  
Articular Chondrocytes ◽  
Soft Agar ◽  
Antiinflammatory Drugs ◽  
Soft Agar Growth ◽  
Rabbit Articular Chondrocytes

A gene for the suppression of anchorage independence is located in rat chromosome 5 bands q22-23, and the rat alpha-interferon locus maps at the same region

1989 ◽  
Vol 92 (2) ◽  
pp. 147-162
Author(s):  
M.Q. Islam ◽  
J. Szpirer ◽  
C. Szpirer ◽  
K. Islam ◽  
J.F. Dasnoy ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Soft Agar ◽  
Chromosome 5 ◽  
Anchorage Independence ◽  
Mouse Hepatoma ◽  
Soft Agar Growth ◽  
Normal Rat ◽  
Rat Fibroblasts ◽  
Interferon Gene

Cell hybrids between malignant mouse hepatoma cells and normal rat fibroblasts with approximately one set of chromosomes from each parent exhibited remarkable karyotypic stability. Most chromosomes of both parents were retained even after prolonged culture in vitro. Normally, such hybrids showed suppression of the transformed phenotype and formed no colonies in soft agar. However, two hybrids, BS140 and BS181, formed a few colonies in soft agar when many cells were seeded, and also occasional foci of cells were detected piling up in monolayer cell cultures. We isolated soft agar colonies (a-subclones) and sub-clones from foci (h-subclones) of both hybrids, and, as a control, subclones of cells from random areas without foci of one hybrid (BS181 p-subclones). When tested for soft agar growth, cells from the a- and h-subclones of both BS140 and BS181 formed colonies at frequencies comparable to the malignant mouse hepatoma parent, whereas the control cells of the BS181 p-subclones (like the normal rat parental cells) yielded no soft agar colonies. All the cell lines were subjected to detailed karyotype analysis in G-banding, which resulted in the finding that cells from the original BS140 hybrid contained at least one copy of each rat chromosome, whereas BS140 a- and h-subclones had lost both copies of rat chromosome 5. Similarly, the original BS181 hybrid contained at least one copy of each rat chromosome, whereas BS181 a- and h-subclones displayed a deletion of the segment q22-23 of rat chromosome 5. In contrast, the control BS181 p-subclones contained one or two copies of non-deleted rat chromosome 5. The conclusion is that a gene for the suppression of anchorage independence is located in the segment 5q22-23. We propose to call this gene SAI1 (for suppression of anchorage independence). Using Southern blotting, we tested whether any of several gene probes, known to correspond to DNA sequences in rat chromosome 5, were homologous to sequences in the deletion. Only one probe, corresponding to the active alpha1-interferon gene, was shown to be located within the deletion. Hence, the SAI1 gene is closely linked to the alpha 1-interferon gene, and might be identical to this locus.

Effect of tumor promoters on soft-agar growth of Swiss 3T3 cells infected with SV40 tsA mutants

1982 ◽  
Vol 3 (7) ◽  
pp. 833-835 ◽  
Author(s):  
L. Daya-Grosjean ◽  
A. Sarasin ◽  
R. Monier
Keyword(s):  
Soft Agar ◽  
Tumor Promoters ◽  
3T3 Cells ◽  
Soft Agar Growth ◽  
Swiss 3T3

scholarly journals Fibronectin receptor overexpression and loss of transformed phenotype in a stable variant of the K562 cell line.

1990 ◽  
Vol 1 (9) ◽  
pp. 637-648 ◽  
Author(s):  
B E Symington
Keyword(s):  
Cell Line ◽  
Solid Phase ◽  
Staining Intensity ◽  
Soft Agar ◽  
Growth Potential ◽  
Cloning Efficiency ◽  
Filamentous Actin ◽  
Fibronectin Receptor ◽  
Soft Agar Growth ◽  
Erythroleukemia Cell

A variant of the K562 erythroleukemia cell line, FA-K562, was selected by cycles of adhesion to solid-phase plasma fibronectin (FN). FA-K562 expresses fourfold more cell-surface alpha 5 beta 1 fibronectin receptor (FNR) than parental K562. In addition to expected differences in adhesion to FN, other differences between FA-K562 and K562 implicate this FNR in the regulation of cell growth and morphology. FA-K562 proliferates slowly in liquid culture, its cloning efficiency in soft agar is only approximately 10% compared with approximately 85% for parental K562, and it is nontumorigenic in nude mice. The reduced soft agar growth potential of FA-K562 involves FNR function, because either glycine-arginine-glycine-aspartate-serine (GRGDS) or monoclonal anti-alpha 5 antibody in the agar medium increased cloning efficiency of FA-K562 about fivefold. Morphologically, FN-adherent FA-K562 become fibroblastoid in appearance, assemble filamentous actin, and differ from K562 in vimentin staining intensity and pattern. Soluble GRGDS peptide inhibits both FA-K562 adhesion to FN and the associated cytoskeletal changes. These findings link the alpha 5 beta 1 FNR to both the transformed phenotype and morphology of FA-K562.

Sign in / Sign up

Export Citation Format

Share Document