1. Side population cells contribute to the genesis of human endometrium
Shunichiro Tsuji, M.D., Momoko Yoshimoto, M.D., Ph.D., Kentaro Takahashi, M.D., Ph.D., Yoichi Noda, M.D., Ph.D., Tatsutoshi Nakahata, M.D., Ph.D., Toshio Heike, M.D., Ph.D. 
Fertility and Sterility 
Volume 90, Issue 4, Pages (October 2008)
DOI: /j.fertnstert
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Isolation of side population cells in human endometrium. Side population (SP) cells were represented by the low Hoechst dye fluorescence at two emission wavelengths, red and blue. (A) Side population cells were clearly detected after Hoechst staining, and gated side population cells account for 0.1% to 1% of the total viable cells. (B) In the presence of verapamil, gated side population cells almost completely disappeared. (C) The proportion of side population cells through the menstrual cycle. Pro.P = proliferative phase, n = 8; Sec.P. = secretory phase, n = 9; ∗ = statistically significant difference.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

3. Figure 2
Characterization of side population (SP) cells in human endometrium by cell surface markers. Fluorescence-activated cell sorter analysis was used to separate side population from non-side population (non-SP) cells. (A) Side population cells were smaller than non-side population cells by forward scatter analysis (FSC). Side population cells expressed the stromal cell marker CD13 (B), the epithelial cell marker EMA (C), the endothelial cell markers CD31 (D) and CD34 (E), and the mesenchymal stem cell markers CD105 (F) and CD146 (G). CD45-negative side population cells were double-stained for CD31 and CD105 (H), and CD31 and CD146 (I).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

4. Figure 3
BCRP1 expression and localization in human endometrium. (A) Reverse transcriptase–PCR analysis indicated that BCRP-1 expression was robust in side population (SP) cells but hardly in non-side population (non-SP) cells. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as an internal control. (B) Immunocytochemistry for BCRP1 (green) and nuclei (blue) in side population (magnification ×200) and non-side population (×100) cells collected by Cytospin centrifuge after FACS sorting. MCF7 cells served as positive controls (×200), and normal mouse IgG antibody was used as a negative control (×200). (D, F, and H) Paraffin-embedded sections of the endometrium were stained with the anti-human BCRP1 antibody. (D and F) BCRP-1 is highly expressed in endothelial cells. (C and E) Isotype control. BCRP-1 in endometrial epithelial cells in the basal layer (H, open arrowhead) was weakly positive compared with the negative control (G, open arrowhead). A small population of epithelial cells in the basal layer (H, filled arrowhead) was as strongly positive as endothelial cells. (G) Isotype control. Note absence of staining in the control sections. V = vessel; M = myometrium. (Original magnification, C and D ×40, E to H ×200.)
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

5. Figure 4
Colony-forming efficiency (CFE). (A) To evaluate growth potential of side population (SP) and non-side population (non-SP) cells, cells were sorted by FACS and grown for 14 days. The number of colonies containing >50 cells (circled) was greater in populations derived from side population than from non-side population cells, on the basis of microscopic examination. (B) Side population cells showed greater colony-forming efficiency than did non-side population cells (P<.05). ∗ = statistically significant difference.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

6. Figure 5
Cell cycle analysis analyzed by Hoechst and pyronin Y staining of side population and non-side population cells. Three individual regions, R1, R2, and R3, correspond with G0, G1, and S/G2+M phases, respectively. (A) Side population cells were sorted directly from the endometrium (G0 85%, G1 14%). (B and C) Side population and non-side population cells were sorted from primary culture. (B) G0 72%, G1 25%. (C) G0 16%, G1 73%.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions

7. Figure 6
In vitro decidualization. Both side population (SP) and non-side population (non-SP) cells undergo decidualization in vitro. (A) Decidualized cells transformed into round cells morphologically with E2 and P treatment. (B) Nondecidualized control cells with E2 alone. (C) Expression of the decidualized markers PRL and IGFBP-1 in side population and non-side population cells by RT-PCR. (D) The concentration of PRL in media did not differ significantly between the two groups. Note that the detection limit of the assay was 0.60 ng/mL.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2008 American Society for Reproductive Medicine Terms and Conditions