1. Enrichment of spermatogonial stem cells from long-term cultured human testicular cells 
Bita Nickkholgh, M.D., Sefika Canan Mizrak, V.M.D., Ph.D., Cindy M. Korver, B.Sc., Saskia K.M. van Daalen, B.Sc., Andreas Meissner, M.D., Sjoerd Repping, Ph.D., Ans M.M. van Pelt, Ph.D. 
Fertility and Sterility 
Volume 102, Issue 2, Pages e5 (August 2014)
DOI: /j.fertnstert
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
Enrichment of spermatogonia in sorted cells. (A) Fold change expression of ITGA6 and GPR125 in various sorted fractions compared with nonsorted cultured testicular cells derived from two patients (two independent cultures for each patient; mean ± SEM). *P≤.05. (B) Relative enrichment of spermatogonia as measured by means of fold change expression (qPCR) in sorted populations compared with nonsorted populations of cultured testicular cells (mean ± SEM) of ZBTB16 (n = 4 independent cultures with sortings for each patient), UCHL1 (n = 4 independent cultures with sortings for each patient) and ID4 (n = 2 independent cultures with sortings for each patient).
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3. Figure 2
COT-1 positive single, paired, and multiple cell human SSC colonies (red, first column) after xenotransplantation of nonsorted cells to immunodeficient mouse testis; DAPI nuclear counterstaining (blue, second column); merged images of colocalization of COT-1 and DAPI (third column), and magnified view of colonies (fourth column). First row shows two COT-1+ colonies: one containing a single cell, and the other one containing multiple cells (four cells); second row shows two COT-1+ colonies containing pairs of cells. The scale bars in the large images and all magnified pictures are 10 μm.
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4. Figure 3
Number of human COT+ SSC colonies in the recipient mouse testes 8 weeks after transplantation of nonsorted, ITGA6 sorted (ITGA6+), HLA and ITGA6 sorted (HLA−/ITGA6+), GPR125 sorted (GPR125+), and HLA and GPR125 sorted (HLA−/GPR125+) cells. Numbers are expressed as the number of colonies per testis per 105 injected cells of the corresponding fraction. (A) Total number of human SSC colonies found in transplanted mice testis. (B) Number of SSC colonies containing single cells (green), pairs (blue), or multiple cells (purple).
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5. Supplemental Figure 1
Histology of testis of both patients (A) URO0034 and (B) URO0077. Both men show normal spermatogenesis.
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6. Supplemental Figure 2
Expression of ID4 in human testis and two independent long-term cultured testicular cells of both included patients. ID4 product size is 144 base pairs (bp). 50 bp ladder was used as an indicator. POLR2A used as reference.
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7. Supplemental Figure 3
ID4 Immunofluorescent staining showing ID4 protein expression (green) of this spermatogonial marker as shown in fine speckles pattern in the nucleus of cells in cytospins of the nonsorted and sorted fractions similar to the transplanted fractions. Columns from left to right: differential interference contrast (DIC) image, ID4 immunofluorencent staining, DAPI, merged pictures. Scale bar: 10 μm.
Fertility and Sterility  , e5DOI: ( /j.fertnstert )
Copyright © 2014 American Society for Reproductive Medicine Terms and Conditions