Image 1_Bi-layered living cellular constructs promote recruitment and activation of an M2 macrophage phenotype.tif

Proteins generated by BLCC and secreted into the media were confirmed and characterized by ELISAs, and this growth media was used as conditioned media to assess the potential recruitment and activation of monocytes/macrophages using THP-1 cells.</p>Results<p>Biopsies of BLCC had outgrowth for up to 9 days with histological characterization confirming presence of a mature epithelium. Genetic assessment identified several genes involved in activation and migration of cells, as well as several genes involved in the recruitment and activation of monocyte/macrophages, including CCL7, G-CSF, and GM-CSF.

Assessment of spent/conditioned media confirmed presence of these proteins and use of spent/conditioned media as a chemoattractant resulted in monocyte recruitment. Cells activated with PMA and exposed to conditioned media were also found to express an M2-like phenotype, as assessed by flow cytometry.</p>Conclusion<p>BLCC wounding promotes monocyte recruitment and subsequent polarization into an M2-like population, a critical immune cell effector for the promotion of a wound healing phenotype.</p>