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C18-4 cell colonies in 3D hydrogels were examined with immunocytochemistry. Cells cultured in 3D hydrogels expressed Vasa but did not express c-Kit. NRGD was varied from 1.5 × 1012 RGD/mm3 (A–C) to 1.3 × 1013 RGD/mm3 (D–F). Cells expressed Vasa at all NRGD, as indicated with arrows (A, D). Cells did not express the c-Kit protein at any NRGD (B, E). (C) and (F) represent negative controls, colonies stained without primary antibodies.
