Figure 3

Characterization of uterine stem cells. A. SCA-1 sorted cells under microscope appear green and are of two distinct sizes including small sized cells which possibly are the very small embryonic-like stem cells VSELs (asterix) and slightly bigger endometrial stem cells EnSCs. B. DAPI stained smear shows that the somatic cells negative for SCA-1 stain nicely with DAPI whereas the stem cells do not stain with DAPI. Similar data was obtained for ovarian stem cells³³; these cells do not stain with DAPI within 2-3 secs possibly because being pluripotent they have open chromatin. C. Higher magnification of OCT-4 stained cells with propidium iodide used as a counterstain shows cells have distinct nuclear staining of OCT-4. D. OCT-4 immuno-localization on the smears. Note the presence of two distinct sized cells which stain positive for OCT-4 including the VSELs (asterix) and slightly bigger EnSCs. E. Nuclear versus cytoplasmic OCT-4 staining can be clearly observed at higher magnification. F. Negative control. Scale bars represent 20 μm. Lower panel G shows the qRT-PCR results of negative (NEG) and positive (POS) fractions after immuno-magnetic sorting of SCA-1 positive cells. As evident the stem cell markers Sca1, Oct 4A and Oct 4 are highly enriched in the positive fraction.