Isolated spermatogonia protrude active pseudopodia in vitro

Area of Science:

• Reproductive Biology
• Cell Biology
• Spermatogenesis

Background:

• Spermatogonia are essential for male fertility as the stem cells of sperm production.
• Distinguishing spermatogonia from other germ cells is crucial for understanding reproductive processes and diseases.

Purpose of the Study:

• To identify a reliable method for identifying spermatogonia in dispersed cell populations.
• To investigate the characteristic features of spermatogonia, including their morphology and behavior.

Main Methods:

• Enzymatic digestion was used to obtain dispersed spermatogenic cells from prepuberal mice, adult male mice, nonazoospermic men, and normospermic men.
• Live cell observation using Normarski optics was performed.
• Immunoreactivity for gamma-H2AX and alkaline phosphatase activity were assessed.

Main Results:

• A distinct subset of cells, identified as spermatogonia, protruded active pseudopodia in all studied groups (47.4% in prepuberal mice, 1.4% in adult mice, 5.1% in nonazoospermic men, 2.4% in normospermic men).
• These pseudopodia-protruding cells exhibited specific morphological characteristics: 8-10 µm diameter, high nucleus-to-cytoplasm ratio, prominent nucleoli, and distinct nuclear membrane.
• Low alkaline phosphatase activity and homogeneous nuclear gamma-H2AX immunoreactivity further supported their identification as spermatogonia.

Conclusions:

• Active pseudopodia protrusion is a reliable indicator for identifying spermatogonia in dispersed cell preparations.
• The observed morphological and molecular features provide a robust signature for spermatogonia, valuable for reproductive research and diagnostics.