116 Chapter 5 RESULTS IPF lung-derived cells form a smaller number of organoids independent of the number of epithelial cells Both IPF and non-IPF lung-derived cell suspensions were able to form organoids by day 7 (Figure 1). We first compared organoid formation efficiency between the IPF and IPF groups in isolated EpCAM+ cell populations. The number of EpCAM+ cells isolated from IPF lung tissue was significantly lower compared to the non-IPF group (Supplementary Figure 1). Therefore, the number of organoids was normalized to the number of epithelial cells isolated per donor to correct for differences in epithelial cell counts. We observed that IPF and non-IPF-derived EpCAM+ cells were equally well capable of forming organoids, as reflected by both the size of the organoids and the numbers (Figure 2A). Next, we assessed organoid formation efficiency of unfractionated suspensions. We normalized for the input of epithelial numbers based on the number of EpCAM+ in the cytospins of these suspensions and observed that significantly more organoids formed from the unfractionated suspensions compared Figure 1: Organoid formation by lung alveolar progenitor cells isolated from IPF and non-IPF lungs. A) non-IPF (unfractionated), B) non-IPF EpCAM+, C) IPF (unfractionated), D) IPF EpCAM+. Brightfield images were taken on day 7 of the organoid cultures with a 2X magnification. Scale bar: 1000 μm. Representative images are shown.
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