An integrin-binding array platform identifies α_vβ₈ and α₅β₁ integrins on rat primary cortical neurons to support...

Integrins are the major cell adhesion receptors expressed in almost all cell types connecting the extracellular matrix with cell cytoskeletons and transducing bi-directional signals across cell membranes. In the central nervous system (CNS), integrins are pivotal for CNS cell migration, differentiation, neurite outgrowth and synaptogenesis in both physiological and pathological conditions. Here we studied the effect of different integrin biding peptides for growth and development of primary cortical neurons in vitro.

Rat primary cortical neurons were cultured in an integrin-binding array platform, which contains immobilized varying short synthetic peptides that bind to 16 individual types of integrin on a 48-well cell culture plate. After cultured for 7 days, cells were fixed and processed for immunostaining with neuronal markers. The overall neuronal growth and neurite outgrowths were quantified.

We found that binding peptides for integrin α_vβ₈, α₅β₁ and α₃β₁ particularly the former two provided superior condition for neuronal growth, survival and maturation. Moreover, optimal neurite outgrowth was observed when neurons were cultured in 3-dimension using injectable hydrogel along with binding peptide for α_vβ₈ or α₅β₁ integrins.

For primary neuronal culture, poly-D-lysine coating is conventional method to support cell attachment. Our study has demonstrated that selected integrin binding peptides provide greater support for the growth of cultured primary neurons.

These data suggest that integrin α_vβ₈ and α₅β₁ are conducive for survival, growth and maturation of primary cortical neurons. This information could be utilized in designing combinational biomaterial and cell-based therapy for neural regeneration following brain injury.