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(A) MiR-204 and miR-211 inhibition of osteogenesis (a), and chondrogenesis (c) coupled with promotion of adipogenesis (b) in human C3 colonic MSC line (as indicated by black arrows for intracellular lipid droplets formation). PGK represent C3 MSCs infected with empty vector viral particles. Human C6 colonic MSC line (insert, i) is shown as negative control being unable to differentiate in cartilage [30]. Scale bar = 30 µm, 4­0 µm. (B) Effect of miR-204 and miR-211 enforced expression in human C3 colonic MSCs hematopoietic supporting activity. Cord blood CD34+ cells were plated onto C3-MSC layers. Weekly cultures were halved to be refed, harvested cells were counted (shown in a) and replated in semisolid hematopoietic secondary cultures to evaluate their progenitor cell content (b). Graphs derive fromone of three different experiments performed in triplicates. C3 represent C3 MSCs infected with empty vector viral particles. BM12-MSCs are included as standard hematopoietic supportive cells. The total number of hematopoietic cells in LTC was unaffected by miR204&211 (a), whereas the number of hematopoietic progenitors (HPCs) was inhibited (b). At wk 5 the culture was terminated harvesting both the cells in suspension (s) and the ones entrapped in the adherent layer (a). Their numbers were added up to generate wk 5 (s+a) points.

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