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. 2018 Mar 22;19(4):943. doi: 10.3390/ijms19040943

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Figure A3 — Application of the 3D culture system for the enrichment of CD133⁺ cells. (a) Schematic of ring colonies formed by pancreatic single cells; (b) representative photograph of the growth of a ring colony taken by inverted fluorescence microscope; (c) representative photograph of ring colonies on the 14th day taken by inverted fluorescence microscope; (d) the proportion of CD133⁺ cells in ring colonies; (e) colony forming frequencies of different passages; (f) the proliferation of pancreatic progenitor-like cells is reflected in the number of cells per well, counted with a cell counter; (g,h) CLSM images of a ring colony, which showed the expressions of Sox9 and Ki67 in the 3D culture system. Nuclei were stained with DAPI; (i) indicates the progenitor-related genes, Sox9, Pdx1, Hnf1b and CD133; differentiation-related genes, ins1, ins2, ngn3 and Pax6; ductal epithelial cells-related genes; CK7; and proliferation-related genes, CyclinD1 and Ki67 in colonies and pancreas; (j) photographs taken by inverted fluorescence microscope of CD133⁺ cells or oligo-treated CD133⁺ cells; (k) colony forming frequencies of CD133⁺ cells and oligo-treated CD133⁺ cells, defined by the number of colonies/the number of plated cells per well. “Relative mRNA levels” are the measured mRNA levels compared to the mRNA levels of the internal standard, cyclophilin A. *** p < 0.001 versus 1.0 g/L colony group; ** p < 0.01 versus 1.0 g/L colony group. * p < 0.05 versus 1.0 g/L colony group. Results represent three independent experiments.

Figure A3 — Application of the 3D culture system for the enrichment of CD133⁺ cells. (a) Schematic of ring colonies formed by pancreatic single cells; (b) representative photograph of the growth of a ring colony taken by inverted fluorescence microscope; (c) representative photograph of ring colonies on the 14th day taken by inverted fluorescence microscope; (d) the proportion of CD133⁺ cells in ring colonies; (e) colony forming frequencies of different passages; (f) the proliferation of pancreatic progenitor-like cells is reflected in the number of cells per well, counted with a cell counter; (g,h) CLSM images of a ring colony, which showed the expressions of Sox9 and Ki67 in the 3D culture system. Nuclei were stained with DAPI; (i) indicates the progenitor-related genes, Sox9, Pdx1, Hnf1b and CD133; differentiation-related genes, ins1, ins2, ngn3 and Pax6; ductal epithelial cells-related genes; CK7; and proliferation-related genes, CyclinD1 and Ki67 in colonies and pancreas; (j) photographs taken by inverted fluorescence microscope of CD133⁺ cells or oligo-treated CD133⁺ cells; (k) colony forming frequencies of CD133⁺ cells and oligo-treated CD133⁺ cells, defined by the number of colonies/the number of plated cells per well. “Relative mRNA levels” are the measured mRNA levels compared to the mRNA levels of the internal standard, cyclophilin A. *** p < 0.001 versus 1.0 g/L colony group; ** p < 0.01 versus 1.0 g/L colony group. * p < 0.05 versus 1.0 g/L colony group. Results represent three independent experiments.