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Studies of gene expression in human cumulus cells indicate pentraxin 3 as a possible marker for oocyte quality Xingqi Zhang, Ph.D., Nadereh Jafari, Ph.D., Randall B. Barnes, M.D., Edmond Confino, M.D., Magdy Milad, M.D., Ralph R. Kazer, M.D. Fertility and Sterility Volume 83, Issue 4, Pages (April 2005) DOI: /j.fertnstert Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions
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FIGURE 1 Profiling gene expression in human cumulus cells. The outer ring represents functional groups of genes that were expressed at levels >1.25-fold higher in cumulus cells from oocytes that developed into 8-cell embryos on day 3 than those from oocytes that failed to fertilize, with a P-value <.01. The inner ring represents functional groups of genes that were expressed at levels >1.25-fold higher in cumulus cells in group A than those in group B. The shaded area denotes genes whose expression was undetectable in the opposing group of cells. Fertility and Sterility , DOI: ( /j.fertnstert ) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions
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FIGURE 2 Verification of the presence of Ptx3 mRNA in human cumulus cells by reverse transcription and polymerase chain reaction (PCR). Total RNA was extracted from human cumulus cells and reverse transcribed to obtain cDNA, which was then subjected to PCR. The amplification products were examined by electrophoresis in 1% agarose. Lane A: total RNA from pooled cumulus cells was directly amplified in PCR using Ptx3 primers (absence of a specific amplification product indicates the absence of a significant amount of genomic DNA contamination); lane B: amplification of cumulus cDNA with Ptx3 primers, with an expected amplification product of 186 bp; lane L: 100-bp ladder. Fertility and Sterility , DOI: ( /j.fertnstert ) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions
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FIGURE 3 Relative abundance of Ptx3 mRNA in human cumulus cells. Total RNA was extracted from individual cumulus complexes and reverse transcribed to obtain cDNA, which was then subjected to real-time polymerase chain reaction (PCR). The relative abundance of Ptx3 mRNA was the difference between the number of cycles of amplification for Ptx3 to reach an arbitrary threshold and the number of cycles of amplification for 18s RNA to reach this threshold in the same sample. The relative abundance of Ptx3 in cumulus cells from fertilized oocytes is expressed as the number of fold changes from that of cumulus cells of unfertilized oocytes. The 8-cell-NI embryos were those in an embryo cohort that were transferred but did not establish a pregnancy; 8-cell-I embryos were those in an embryo cohort that were transferred and that led to the establishment of a clinical pregnancy. *Compared with cumulus cells from unfertilized oocytes, P<.01 (t-test). Fertility and Sterility , DOI: ( /j.fertnstert ) Copyright © 2005 American Society for Reproductive Medicine Terms and Conditions
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