Screening of Integrin Heterodimers Expressed Functionally on the Undifferentiated Spermatogonial Stem Cells in the Outbred ICR Mice

Outbred mice are widely used in toxicology, pharmacology, and fundamental biomedical research. However, there have been no reports of in vitro culture systems for spermatogonial stem cells (SSCs) derived from these mice.

As a step towards constructing a non-cellular niche supporting the in vitro maintenance of outbred mouse SSC self-renewal, we systematically investigated the types of integrin heterodimers that are expressed transcriptionally, translationally, and functionally in SSCs derived from Imprinting Control Region (ICR) mice.

Among the genes encoding 25 integrin subunits, integrin α₁, α₅, α₆, α₉, α_V, and α_E, and integrin β₁ and β₅ had significantly higher transcriptional levels than the other subunits. Furthermore, at the translational level, integrin α₅, α₆, α₉, α_V, α_E, and β₁ were localized on the surface of SSCs, but integrin α₁ and β₅ not. Moreover, significantly stronger translational expression than integrin α₉ and α_E was observed in integrin α₅, α₆, α_V, and β₁. SSCs showed significantly increased adhesion to fibronectin, laminin, tenascin C and vitronectin, and functional blocking of integrin α₅β₁, α₆β₁, α₉β₁ or α_Vβ₁ significantly inhibited adhesion to these molecules.

We confirmed that integrin α₅β₁, α₆β₁, α₉β₁ and α_Vβ₁ actively function on the surface of undifferentiated SSCs derived from outbred ICR mice.