After removal of the cells, the collected buffer was used as conditioned medium.

Total volumes were 150μl for lysate samples and 7ml for conditioned medium samples.

Analysis of the conditioned medium from Ax4 cells indicted that extracellular Countin increased with time as was expected fig.

Most (87%) of the mitogenic activity expressed in GGFHBS5 transfections was present in the conditioned medium.

In contrast, we did not detect significant amounts (over mock transfected control samples) of activity in conditioned medium from the GGFHFB1 transfection.

The first in vitro differentiation and growth of a pure population of mouse mast cells has been carried out using conditioned medium derived from concanavalin A-stimulated splenocytes.

For detecting Countin at each time point, 20 μl of conditioned medium or 10 μg of protein from the cell lysates was fractionated by PAGE.

The mitogenic activity of the GGFHFB1-encoded protein was detected within lysates derived from transfected cells and was not detected in conditioned medium.

Here, only recombinant human GGF-II (from GGFHBS5) is secreted at significant levels into conditioned medium.

We transiently expressed several cDNAs in mammalian cells, then assayed both cell lysates and conditioned medium in a Schwann cell proliferation assay.

In contrast, Egfl7 produced in conditioned medium is a chemo-attractant for rat vascular smooth muscle cells, mouse endothelial cells and for primary mouse embryonic fibroblasts in vitro.

Abbreviations eIF2α, eukaryotic initiation factor 2 alpha; CMF, conditioned medium factor; CM, conditioned medium.

In contrast to the other heregulins, biologically active (p185 erbB2 receptor tyrosine kinase activation) heregulin β 3 is not detected in conditioned medium from transiently expressing COS cells.

Comparison of reactions in the absence and presence of CM showed that the specific PEDF-binding to Y-79 cell-surface receptors was 6.8-fold higher with conditioned medium than with defined medium (Fig.

Co-transfection and parallel transfection experiments using a non-secreted marker protein (β -galactosidase plasmid) confirmed that GGF activity in conditioned medium results from secretion and not simply from cell lysis or leakage.

To stimulate the NCCIT cells with Wnt, we used tissue culture medium containing active Wnt-3A protein produced by mouse L cells (Wnt-3A CM [ 8 ] ), in comparison to control conditioned medium (CCM).

A large protein complex, counting factor, was purified from conditioned medium and shown to be involved in causing streaming cells to break up into groups of cells in order to generate mounds and subsequent developmental structures of the desired size [ 12 13 ] .

Development of the cells in monolayers in buffer or in conditioned medium from BS153 or Ax4 cells [ 10 ] indicated that secreted factors were involved in causing both the earlier aggregation of BS153 cells and the larger size of the resulting multicellular entities.

To investigate this ability of different recombinant GGFs to be secreted, we determined for the two human cDNA clones the distribution of total expressed GGF (Schwann cell mitogenic) activity between cell lysate and conditioned medium after transient mammalian transfections (Table 2).

Takigawa M, Shirai E, Enomoto M, et al.: A factor in conditioned medium of rabbit costal chondrocytes inhibits the proliferation of cultured endothelial cells and angiogenesis induced by B16 melanoma: its relation with cartilage-derived anti-tumor factor (CATF).