Cell plating density alters the ratio of estrogenic to progestagenic enzyme gene expression in cultured granulosa cells  Valério M. Portela, D.V.M., Ph.D., Gustavo Zamberlam, D.V.M., M.Sc., Christopher A. Price, Ph.D.  Fertility and Sterility  Volume 93, Issue 6, Pages 2050-2055 (April 2010) DOI: 10.1016/j.fertnstert.2009.01.151 Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Effect of cell plating density on E2 and P secretion, and the E2:P ratio from bovine granulosa cells in long-term serum-free culture. Data are mean ± SEM of three replicate cultures. Bars with different letters are significantly different (P<.05). Fertility and Sterility 2010 93, 2050-2055DOI: (10.1016/j.fertnstert.2009.01.151) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Effect of plating density on abundance of mRNA encoding (A) estrogenic enzymes and FSH receptor and (B) progestagenic enzymes and StAR. Data generated by real-time polymerase chain reaction are expressed relative to a calibrator sample by the ΔΔCt method and are presented as mean ± SEM of three replicate cultures. Bars with different letters are significantly different (P<.05). Fertility and Sterility 2010 93, 2050-2055DOI: (10.1016/j.fertnstert.2009.01.151) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Effect of plating density on the proportion of dead cells and abundance of mRNA encoding the cell damage–inducible gene GADD45B and the cell proliferation gene CCDN2. Cell survival data were generated by flow cytometry, and mRNA abundance was measured by real-time polymerase chain reaction. Data are presented as mean ± SEM of three replicate cultures. Bars with different letters are significantly different (P<.05). Fertility and Sterility 2010 93, 2050-2055DOI: (10.1016/j.fertnstert.2009.01.151) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions