Reconstituted basement membrane reduces proliferation and increases prolactin expression of GH3 cells

This study investigated the effects of reconstituted basement membrane Matrigel on the proliferation and prolactin expression of GH3 cells in culture for 6 days. When cells were cultured on Matrigel, the initial attachment was increased but the cell number was not changed with time whereas rapid increase in cell number was observed in cultures on plastic. Bromodeoxyuridine (BrdU)-labeling showed that BrdU incorporation ratio of GH3 cells cultured on Matrigel was about one half of that observed with cells cultured on plastic (9.7+/-0.7% vs. 18.7+/-1.2%). Immunocytochemistry revealed that the ratio of the prolactin-immunoreactive GH3 cells was about 3.6 times (58.4+/-2.9% on Matrigel vs. 16.2+/-1.4% on plastic), which was compatible with the results of Western blot analysis. In situ hybridization demonstrated that prolactin mRNA-positive cells were identified more frequently when cells were cultured on Matrigel compared to cultures on plastic. These findings indicate that Matrigel is a proper culture substrate for the long-term culture of GH3 pituitary cells due to the inhibition of overgrowth and promotion of prolactin expression.