Chemokine (C-X-C) Ligand 12 Facilitates Trafficking of Donor Spermatogonial Stem Cells

Figure 2

Effects of CXCR4 inhibitor AMD3100 on SSC-enriched THY1⁺ mouse germ cells. (a) An equal number of SSC-enriched cultured germ cells (1 × 10⁵ cells/well) were cultured on a layer of STO feeder cells and were treated with the vehicle (DMSO) or treated with AMD3100 (1.25 μM), CXCL12 (10 ng/mL), or the combination of both CXCL12 and AMD3100. After 7 days the total number of cultured germ cells was quantified by cell counting. Values are representative of three independent experiments, and significance was calculated using one-way ANOVA. Data are means ± SEM. (b) Impact of AMD3100 on germ cell viability. SSC-enriched cultured germ cells were seeded on feeder-free, laminin-coated 96-well plates at a density of 1 × 10⁴ cells/well. Cells were either untreated (WT) or treated with DMSO control, AMD3100 (1.25 μM and 6.25 μM), or AKT inhibitor, LY29004 (33 μM). Cell viability was assessed at days 4 and 6 following treatment using the ATPLite assay. Data are means ± SEM, where significance (denoted as ) was calculated using Student’s -test (, ).

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