A novel protocol for isolation and culture of multipotent progenitor cells from human urine

Cell therapy holds promise for treating a variety of diseases. Seeking available source of adult stem cells remains a great challenge in cell therapy. Urine is considered as an ideal source of adult stem cells which can be easily acquired by noninvasive methods. However, specific cell types in urine have not been well documented. Here, the aim of our study is to identify cell types in urine, and isolate and expand progenitor/stem cells from human urine and further evaluate their multipotency. Urine samples were collected from healthy donors. The cell suspension was seeded and selected because of plastic adherence. Colonies with two different morphologies appeared 7 days later. One type of colony was spindle-shaped and fibroblast-like; the other cell type displayed rounder shape. Cells that displayed fibroblast-like shape were selectively enriched using a cloning cylinder. Then multidifferentiation induction assays and immunophenotyping assays were applied. Characterization assays indicated that adherent cells possessed potent trilineage differentiation capacity and expressed CXCR4 and Nanog, as well as some mesenchymal stem cell surface antigens (including CD90 and CD44). Taken together, at least two cell populations exist in human urine. A stem cell subpopulation with trilineage differentiation capacity from human urine can be selectively enriched using the cloning cylinder method. Urine may become an ideal source of adult stem cells for cell therapy and further clinical implications.