Jekyll or Hyde: does Matrigel provide a more or less physiological environment in mammary repopulating assays? - PubMed (original) (raw)

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

François Vaillant et al. Breast Cancer Res. 2011.

Abstract

In vivo transplantation is the current 'gold-standard' assay for evaluating mammary stem cell (MaSC) function. Matrigel, a reconstituted extracellular matrix derived from a mouse sarcoma line, is increasingly being utilized for mammary repopulating assays, although original studies were carried out in its absence. This matrix has also been shown to enhance tumor-initiating capacity. Whilst Matrigel increases the rate of engraftment by MaSCs, it also appears to promote progenitor activity that is distinct from bona fide stem cell activity. This caveat should be considered when interpreting mammary reconstitution assays that incorporate Matrigel, particularly when transplanting high cell numbers.

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Figures

Figure 1

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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