Figure 1
Quantitative and qualitative analysis of ALDH positive MECs in virgin and parous glands. Mammosphere formation assay (n = 6) was established from virgin and parous MECs after cell dissociation of the mammary gland. A) Mammospheres obtained from virgin and parous rat MECs after 7 days in culture B) Percentage of ALDHbright virgin and parous MECs using flow cytometry. The positively stained cells were detected using the green fluorescence channel (520–540 nm) of the flow cytometer. C) Mammosphere formation efficiency; total number of mammospheres formed after 7 days in culture were counted and divided by the total number cells initially plated. D) Immunofluorescence images showing the presence of stem cell markers like SOX2 and OCT4 in mammospheres obtained from MECs of both the groups. Mammospheres, after 7 days in culture were placed on poly-l-lysine coated chamber slides for attachment for 3-4 hrs and followed by sequential double immunofluorescence staining procedure. The cells from both groups showed positivity for SOX2 and OCT4 indicating the enrichment of stem cells in the mammospheres.