#hMADS adipocytes and hMADS preadipocytes do not alter trastuzumab. (A) Binding of fluorescein isothiocyanate (FITC)-conjugated trastuzumab to BT-474, differentiated human multipotent adipose-derived stem cells (#hMADS) and human multipotent adipose-derived stem cells (hMADS). Dotted red lines indicate unstained cells, and solid green lines indicate cells labeled with FITC-conjugated trastuzumab. (B) Antibody-dependent cellular cytotoxicity (ADCC) assays performed with 1 μg/ml or 10 μg/ml trastuzumab in the presence of #hMADS or hMADS. (C) Binding of FITC-conjugated trastuzumab, previously incubated in #hMADS-CM or hMADS-CM or in the control media, on BT-474 cells (1 μg/ml) or NK-92-CD16 cells (10 μg/ml). Dotted red lines indicate unstained cells, dashed green lines indicate FITC-conjugated trastuzumab preincubated in the control media and solid blue lines indicate FITC-conjugated trastuzumab preincubated in conditioned media (CM) from hMADS or hMADS (#hMADS-CM or hMADS-CM, respectively). (D) ADCC assays performed with fresh trastuzumab or trastuzumab preincubated overnight in the control media or in #hMADS-CM (left) or hMADS-CM (right). Results representative of three independent experiments are shown in (A) and (C). Mean ± SD values of three independent experiments, each performed in triplicate, are shown in (B) and (D). *P < 0.05; **P < 0.01.