{Reference Type}: Journal Article
{Title}: Myogenic commitment of human stem cells by myoblasts Co-culture: a static vs. a dynamic approach.
{Author}: Scala P;Lovecchio J;Lamparelli EP;Vitolo R;Giudice V;Giordano E;Selleri C;Rehak L;Maffulli N;Della Porta G;
{Journal}: Artif Cells Nanomed Biotechnol
{Volume}: 50
{Issue}: 1
{Year}: Dec 2022
{Factor}: 6.355
{DOI}: 10.1080/21691401.2022.2039684
{Abstract}: An in-vitro model of human bone marrow mesenchymal stem cells (hBM-MSCs) myogenic commitment by synergic effect of a differentiation media coupled with human primary skeletal myoblasts (hSkMs) co-culture was developed adopting both conventional static co-seeding and perfused culture systems. Static co-seeding provided a notable outcome in terms of gene expression with a significant increase of Desmin (141-fold) and Myosin heavy chain II (MYH2, 32-fold) at day 21, clearly detected also by semi-quantitative immunofluorescence. Under perfusion conditions, myogenic induction ability of hSkMs on hBM-MSCs was exerted by paracrine effect with an excellent gene overexpression and immunofluorescence detection of MYH2 protein; furthermore, due to the dynamic cell culture in separate wells, western blot data were acquired confirming a successful cell commitment at day 14. A significant increase of anti-inflammatory cytokine gene expression, including IL-10 and IL-4 (15-fold and 11-fold, respectively) at day 14, with respect to the pro-inflammatory cytokines IL-12A (7-fold at day 21) and IL-1β (1.4-fold at day 7) was also detected during dynamic culture, confirming the immunomodulatory activity of hBM-MSCs along with commitment events. The present study opens interesting perspectives on the use of dynamic culture based on perfusion as a versatile tool to study myogenic events and paracrine cross-talk compared to the simple co-seeding static culture.