Deleted in Azoospermia-Like (DAZL) gene–expressing cells in human amniotic fluid: a new source for germ cells research? Konstantinos Stefanidis, M.D.,

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Deleted in Azoospermia-Like (DAZL) gene–expressing cells in human amniotic fluid: a new source for germ cells research? Konstantinos Stefanidis, M.D., Dimitris Loutradis, M.D., Lemonika Koumbi, M.Sc., Vasiliki Anastasiadou, M.D., Vasiliki Dinopoulou, B.Sc., Erasmia Kiapekou, M.D., Alexandros A. Lavdas, Ph.D., Spiros Mesogitis, M.D., Aris Antsaklis, M.D. Fertility and Sterility Volume 90, Issue 3, Pages 798-804 (September 2008) DOI: 10.1016/j.fertnstert.2007.06.106 Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Cells in human amniotic fluid express DAZL (A) and Oct-4 (B) mRNA. Reverse transcriptase polymerase chain reaction analyses of DAZL and Oct-4 mRNA expression in a representative sample of amniotic fluid cells containing DAZL and Oct-4-positive cells (FAFC), with (+) and without (−) reverse transcriptase. Flow cytometric analysis of amniotic fluid cells of 7AAD, DAZL, and C-kit. Amniotic fluid cells were identified by two scatter regions, R1 and R2, on an FSC vs. SSC dot plot (C), and only the 7AAD-negative cell populations, R3 and R4, corresponding to R1 and R2 populations, respectively, were further analyzed (gating was set by the unstained sample) (D, E). The overlay graphs show the viable cell populations selected from gate R1 and R3 for DAZL and C-kit; green stripes represent the expression levels of DAZL (F) and C-kit (G) and the red stripes their corresponding isotype controls. Fertility and Sterility 2008 90, 798-804DOI: (10.1016/j.fertnstert.2007.06.106) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Amniotic fluid cells after 1 day in vitro immunostained for Oct 3/4 (A, B), SSEA-4 (D, E), DAZL (G, H), and vimentin (J, K) as seen under the confocal microscope. To-Pro-3 was used to stain nuclei (A, D, G, J). Phase contrast in C, F, I, and L. Scale bars = 40 μm (A–F, J–L) and 20 μm (G–I). Fertility and Sterility 2008 90, 798-804DOI: (10.1016/j.fertnstert.2007.06.106) Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions