Effect of OSB replenishment on long-term ex vivo maintenance of MMC.
(a) OSB cell viability in coculture was assessed by co-staining cultures with CD13 and 7-AAD to quantify dead OSB by flow cytometric analysis. ‘OSB alone’ bars are shown as reference. (b) MMC viability was evaluated by counting live and dead MMC cells (as determined by morphological analysis using Giemsa staining). Data presented is an average count of 18 separate fields. * p < 0.05, ** p < 0.01. (c-f) Giemsa staining of 4 weeks old cultures of OSB alone or OSB + BMMC coculture with (c-d) OSB refilled after 2 weeks, and (e-f) no replenishment.