118 Chapter 5 Cell fractions in unfractionated lung suspensions do not differ significantly between IPF and non-IPF groups To investigate whether there were differences in the composition of cell types in the unfractionated suspensions between IPF and non-IPF lungs, cytospins of unfractionated suspensions were stained for several markers. As the stromal compartment of the lungs includes various major cell types, we stained for epithelial cells (EpCAM), stromal cells (CD90), macrophages (CD68) and endothelial cells (CD31), which have all been implicated in IPF pathology [3, 9]. We were able to identify CD90+, CD68+ and CD31+ cells in the unfractionated suspensions, but we did not observe significant differences in the percentages of each type of cell between the non- IPF and IPF groups (Figure 3). Figure 3: Percentage of cells stained positive for EpCAM (epithelial cells), CD31 (endothelial cells), CD90 (mesenchymal cells) and CD68 (macrophages) in unfractionated cell suspension cytospins from IPF and non-IPF lungs. Means ± SD values are shown. Statistical differences between IPF and non-IPF groups were tested using the unpaired t-test after verifying data normality with Q-Q plots and Shapiro-Wilk test. DISCUSSION In this study, we compared the organoid forming efficiency of EpCAM+ epithelial cell populations and unfractionated cell suspensions from parenchymal regions of IPF and non-IPF lungs. Our results suggest the presence of a supportive cell population in the unfractionated lung suspensions. Of interest, we observed reduced organoid forming efficiency in the unfractionated suspensions from IPF compared to non-IPF lungs. This difference was not present in organoids formed from isolated EpCAM+ cell populations from both IPF and non-IPF lungs, suggesting that dysregulated cross-
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