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Figure 1 | Breast Cancer Research

Figure 1

From: Jekyll or Hyde: does Matrigel provide a more or less physiological environment in mammary repopulating assays?

Figure 1

Effect of Matrigel on the transplantation of mammary epithelial cell subpopulations. (a) Table showing the number of outgrowths per number of mammary fad pads injected with either 75 CD29hiCD24+ (mammary stem cell (MaSC)-enriched) cells or 1,000 CD29loCD24+ (luminal) cells, in either 0%, 25% or 50% Matrigel. Single cell suspensions were prepared from the mammary glands of 8-week-old to 10-week-old FVB/N-Rosa26 female mice, labeled with fluorochrome-conjugated antibodies and double-sorted as described [2]. The MaSC-enriched and luminal cell populations were identified following depletion of endothelial and hematopoietic cells using anti-CD45, anti-CD31 and anti-TER119 antibodies. Cells were injected (10 μl volume) into the cleared inguinal mammary fat pads of 3-week-old FVB/N female recipients and were collected 8 weeks post transplantation for X-gal staining. β-Gal+ branched ductal structures were scored as positive. Data are shown for four independent experiments. (b) Images of X-gal-stained outgrowths: outgrowth derived from transplantation of 75 CD29hiCD24+ cells in 50% Matrigel (top), and largest outgrowth obtained from transplantation of 1,000 CD29loCD24+ cells in 50% Matrigel (bottom). Bar = 1 mm. (c) Bar chart representation of mammary outgrowths as a function of fat-pad filling following transplantation of each subpopulation. The axes shown differ for the two populations, since very few structures were generated by the CD29loCD24+ population and these did not exceed 5%. Data are shown for four independent experiments. MFP, mammary fat pad.

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