Fig. 1. Xenotransplantation of DCCs.

a Diagnostic bone marrow aspirates from breast (BrCa, n = 19) or prostate (PC, n = 27) cancer patients (M0- or M1-stage of disease) were either CD45-depleted, enriched for EpCAM, or cultured under sphere conditions. Resulting spheres, CD45-depleted, or EpCAM-enriched BM cells were injected intra-venously (i.v.), intra-femorally (i.f.), sub-cutaneously (s.c.), sub-renally (s.r.), or into the mammary fat pad (mfp) of NOD-scid or NOD-scidIL2Rγ-/- mice. Mice with sub-cutaneous or mammary fat pad injections were palpated weekly. All other mice were observed until signs of illness or were sacrificed after 9 months. Injection routes that led to xenograft formation are highlighted in red. b Immunohistochemistry for estrogen-receptor (ER), progesterone-receptor (PR), prostate-specific antigen (PSA), Ki-67, or H & E staining of M1-DCC-derived xenografts is shown. c Human EpCAM- or cytokeratin 8/18/19-expressing DCCs were detected in the BM of 4/42 mice transplanted with M0-stage patient samples. DCCs from two of the four mice were isolated and their human origin was verified by a PCR specific for human KRT19. Pure mouse or human DNA was used as control. 1, 2 = cytokeratin 8/18/19-positive DCCs; N = cytokeratin 8/18/19-negative BM-cell, P = pool of BM-cells of recipient mouse; m = mouse positive control; h = human positive control, c = non-template control. d Single cell CNA analysis of the EpCAM-expressing DCC isolated at 4 weeks after injection from NSG BM (c) and a human hematopoietic cell as control. Red or blue indicate gain or loss of chromosomal regions.