Figure 1.

Establishment and characterization of single cell-derived colonies from human kidney. (A) CFE of single cell clones derived from fresh hAK cells according to their size (CFE% = 10.29 ± 1.1); Small (S)% = 4.69, Medium (M)% = 2.47, Large (L)% = 3.125; (B) Representative graph of self-renewal capacity of hAK-clones base on colony size. Data are presented for each passage as relative number of clones generated from the total number of cells plated. Most colonies surviving expansion were large (L) colonies (100% at P1 and 33.33% continued to expand for up to 4 passages) compared to medium (M) colonies that presented limited ability for clonal expansion (63% at P1 and none continued to expand beyond the 3rd passage) and small (S) colonies that failed to survive along passages altogether; (C) Representative graph of the expansion potential of hAK single cell derived clones. Clones originating from a single cultured AK cell were able to expand into approximately 3.4 * 10⁶ cells; (D) Representative morphology of an expanded hAK single cell derived clone (D6-1) along passages, compared to the heterogeneous hAK culture from which the clones was derived. A stable epithelial (EL) phenotype was preserved during clonal expansion for several month in contrast to the heterogeneous pool of cultured cells, which were already undergoing senescence and switching to a fibroblast-like morphology at the same time point. Scale bars, 100 μm; (E) Representative images of each of the 9 single cell clones at P0; (F) Representative photomicrographs of single cell clonal phenotypes. Two type of clones were generated: Epithelial-like (ELC) or Fibroblast-like (FLC). Scale bars, 100 μm.