Developmental Changes in the in Vitro Activated Regenerative Activity of Primitive Mammary Epithelial Cells (original) (raw)
Figure 3
Production of MRUs and CFCs in 7-d Matrigel cultures of adult and fetal mammary cells.
(A) General experimental design. Mammary cells were added to 50 µl of solidified Matrigel (±2.5×104 irradiated 3T3 fibroblasts) in 200 µl of medium and incubated for 7-d. Each well was then examined for the presence of one or more visible structures and the contents then fixed and stained or dissociated into a suspension of viable single cells to perform FACS, CFC, or MRU assays for comparison with corresponding starting (input) values. (B) CFC outputs from 7-d Matrigel cultures of unseparated adult mammary cells as a function of the input cell number (expressed as the number of input EpCAM+ cells) and the addition of irradiated fibroblasts. (C) CFC outputs from 7-d cultures of unseparated fetal cells as a function of the input cell number (expressed as the number of input EpCAM+ cells). Comparison of the corresponding relationship for adult cells (dotted line redrawn from (B)) shows a ∼5-fold higher CFC output by the fetal cells. (D) Comparison of increased numbers of MRUs obtained from 7-d Matrigel cultures of fetal and adult mammary cells (values determined by LDA as described in Tables S1, S2, S3, S4, S5, S6 and expressed relative to 100 EpCAM+ cells).