Suppressed OSB activity and viability, resulting in the loss of BMMC/MMC long-term viability.
(a) Relative alkaline phosphatase (ALP) activity (a.u.) of OSB at Day 0, 5 and 10 of coculture. Taken into consideration the 4 day of OSB alone culture prior to BMMC seeding (in OSB culture medium), the actual culture time for OSB was 4, 9 and 14 as described in the methods section. (b) At each time point, OSB cell viability in coculture was assessed by co-staining cocultures with CD13 (an OSB marker) and 7-AAD to quantify dead OSB by flow cytometric analysis. ‘OSB alone’ bar is shown as reference. (c) Primary patient-derived BMMC were labeled with CFSE prior to coculture. BMMC cell viability was assessed by co-staining cultures with 7-AAD to quantify dead BMMC by flow cytometric analysis at indicated coculture time. * p < 0.05, ** p < 0.001.