Journal: F&S science

Article Title: Blastocyst development after fertilization with in vitro spermatids derived from nonhuman primate embryonic stem cells

doi: 10.1016/j.xfss.2021.09.001

Figure Lengend Snippet: Nonhuman primate embryonic stem cell (nhpESC) histone 2B-green fluorescent protein (H2B-GFP) cells differentiated in spermatogonial stem cell (SSC) conditions yield haploid, round spermatid-like cells. ( A ) Representative cell cycle flow cytometry profile data from one of five separate differentiations (n = 5) ( left image ) of nhpESC H2B-GFP cells differentiated in SSC conditions for 10 days. The dark green peak on the left-hand side of the graph represents the 1N peak ( asterisk ). ( B ) Fluorescence in situ hybridization (FISH) confirms haploidy. Representative FISH images from five different (n = 5) nhpESC H2B-GFP cells differentiated for 10 days in SSC conditions. The FISH Probe for chromosome 1 ( green ) shows a diploid cell ( left image ) with two chromosome 1s ( arrow indicating two green dots ), and sorted haploid cells ( right image ) show one chromosome 1 (arrow indicating one green dot ). Bar = 5 μ m. ( C ) Following our 10-day differentiation, haploid cells were sorted by fluorescence-activated cell sorting and immunostained with antibodies against postmeiotic spermatid markers ( red , third column ) acrosin ( top row ), Protamine 1 (PRM1; middle row ), and Transition Protein 1 (TNP1; bottom row ). H2B-GFP ( green , second column ); Hoechst stain ( blue , first column ) for deoxyribonucleic acid. Merge of fluorescence images ( fourth column ). Bar = 10 μ m. Data shown are representative of five separate differentiations (n = 5). ( D ) Representative images of round spermatids from a rhesus testis cell biopsy ( left ) and an in vitro-derived round spermatid-like cell ( right ) stained with 4′,6-diamidino-2-phenylindole ( blue ), Protamine 1 ( red ), and acetylated tubulin ( green ). Bar = 5 μ m.

Article Snippet: Acrosin (ACR), Protamine 1 (PRM1), and Transition Protein 1 were from Santa Cruz Biotechnology.

Techniques: Flow Cytometry, Fluorescence, In Situ Hybridization, FACS, Staining, In Vitro, Derivative Assay

Journal: Asian journal of andrology

Article Title: Characterization of nucleohistone and nucleoprotamine components in the mature human sperm nucleus

doi: 10.1111/j.1745-7262.2008.00410.x

Figure Lengend Snippet: Table 1

Article Snippet: Mouse monoclonal anti-human protamine 1 (PRM1) and protamine 2 (PRM2) antibodies were a generous gift from Dr Rod Balhorn, Lawrence Livermore National Laboratory (Livermore, CA, USA).

Techniques: Immunofluorescence