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. 2015 Nov 5;5(6):979–987. doi: 10.1016/j.stemcr.2015.10.003

Figure 2.

Figure 2

Comparison of PTK7 to Other Stem Cell Markers Proofs Enhanced Capacity to Enrich for Self-Renewing Organoid Cells

(A) Human colonic organoids maintained in WREN media were disaggregated and stained against the indicated surface markers. Via FACS, 9%–10% of cells showing the brightest staining intensity (HIGH) and 20%–21% of cells showing the weakest intensity (LOW) were isolated. Control (transparent profile) and specific staining (gray profile) are shown in each histogram.

See Figures S2A–S2C for an equivalent experiment derived from a different individual.

(B) Organoid formation capacity of cellular fractions isolated via FACS as shown in (A). Each cellular fraction was seeded four times (biological replicate), and the frequency of organoid formation was assessed 8 days after seeding. Error bars indicate SEM. Significance of PTK7 high versus low enrichment over EPHB2, CD44, or CD133 performance was assessed using the F-test (Supplemental Experimental Procedures). ∗p < 0.02.

(C) qRT-PCR analysis of stem cell and differentiation marker gene expression on organoid cells HIGH and LOW for the indicated marker surface abundance according to (A) (see also Figure S2). Gene expression was assessed three times (technical replicate), and error bars indicate SD.