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. 2019 Jun 21;14(6):e0218194. doi: 10.1371/journal.pone.0218194

Fig 1. Study design demonstrating three-step experimental scheme.

Fig 1

Step 1: Characterization of macaque testicular markers by mRNA and protein analysis. Step 2: Spermatogonial isolation, separation of supernatant (SN) and adherent (AT) fractions and sub-culture in three media conditions (Spermatogonial stem cell (SSC), Stemmacs (SM) and Pluristem (PS) media). Gene expression analysis of cultured cells collected at day 7, 14 and 21. Step 3: Seeding of isolated spermatogonia (25,000 cells per well) in 8-well chamber slides with three media conditions. Fixation of cells cultured in three media conditions in 4% Para-formaldehyde (PFA) at day 7, 14 and 21. Immunofluorescence analysis for expression of germ cell markers MAGEA4 and VASA in vitro and quantification of cells and clusters expressing MAGEA4 in vitro during culture.