A The renal papilla is the stem cell niche in the kidney, defined by region-specific cues in the extracellular matrix of the papilla. B The renal cortex, medulla, and papilla were dissected and decellularized separately to obtain kidney region-specific ECM. C ECM was prepared in sheets, hydrogels, and solubilized forms for cultivation of two types of stem cells (kidney stem cells and mesenchymal stem cells). To obtain kidney region-specific ECM sheets, kidney regions were dissected before decellularization or the regional matrix was punched from decellularized whole kidney sections. Alternatively, pre-sectioned regions were decellularized, snap-frozen in liquid nitrogen, ground into a fine powder, lyophilized, and pepsin-digested to yield kidney region-specific ECM digests that were neutralized to obtain solubilized ECM forms. By treating these digests with salt, base, and heat, we obtained kidney region-specific ECM hydrogels. Kidney stem cells (KSCs) that are native to the papilla were cultured on ECM sheets, hydrogels, or solubilized forms derived from the three kidney regions (cortex, medulla, papilla) and compared to mesenchymal cells (MSCs) cultured under the same conditions.