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. Author manuscript; available in PMC: 2016 May 1.
Published in final edited form as: Nat Biotechnol. 2015 Nov;33(11):1193–1200. doi: 10.1038/nbt.3392

Figure 3. Induction of pre-tubular aggregates and renal vesicles from nephron progenitor cells.

Figure 3

(a) Diagram of differentiation into renal vesicles. (b) Whole-well scan for LHX1 in 24-well on day 14 of differentiation. The combination of FGF9 10 ng/ml and transient CHIR 3 µM treatment enhanced LHX1 expression. n=2. Scale bar: 5 mm. (c) Representative images of brightfield and immunocytochemistry for PAX8 and LHX1 in hESCs on day 14. Scale bar: 100 µm. (d) Flow cytometry for PAX8 and LHX1 in hESCs on day 14. Samples treated with secondary antibodies alone were used as controls (gray). (e) Immunocytochemistry for BRN1, HNF1β and LAM (Laminin) on day 14 of differentiation. n=6. 50 µm (the lower panel). (f) Brightfield imaging of the organoids that formed in culture after cells were resuspended on day 9, transferred to ultra-low attachment 96-well plates and studied on day 14. Controls were cultured in the basic differentiation medium (ARPMI) after resuspension. FGF9 and CHIR increased the size of the organoids. Scale bar: 100 µm. (g) Whole-mount staining of the organoids on day 11 and 14. Polarized structures surrounded by Laminin were found on day 14, suggesting the differentiation into renal vesicles. n=2. Scale bars: 100 µm.