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1.
Figure 6

Figure 6. From: Prospective isolation and characterization of committed and multipotent progenitors from immortalized mouse mammary epithelial cells with morphogenic potential.

Hypothetical model of stem cell differentiation during mammary gland development. Mammary gland development begins by asymmetric self-renewal in a stem cell, which generates multipotent and bipotent ductal and alveolar progenitor cells. Ductal and alveolar progenitor cells are bipotent, express high CK5 levels and may give rise to luminal and myoepithelium-restricted progenitors. Upon commitment to transient amplification and differentiation, restricted progenitor cells may downregulate CK5 expression.

Frances S Kittrell, et al. Breast Cancer Res. 2011;13(2):R41-R41.
2.
Figure 2

Figure 2. From: Prospective isolation and characterization of committed and multipotent progenitors from immortalized mouse mammary epithelial cells with morphogenic potential.

Hematoxylin and eosin (H&E) staining of alveolar progenitor clone makes lipid droplets indicative of alveolar differentiation. (A) Alveolar progenitor-derived outgrowths following hormonal stimulation using pituitary isografts show the formation of lipid droplets inside the lumen. (B) Ductal progenitor outgrowths, shown for comparison, are incapable of alveolar differentiation, and the ducts lack the ability to form lipid droplets. (C to E) Intraductal papillary hyperplasia (IDH) generated by the ductal progenitors. Arrows indicate IDH lesions. H&E images taken at ×20 original magnification.

Frances S Kittrell, et al. Breast Cancer Res. 2011;13(2):R41-R41.
3.
Figure 4

Figure 4. From: Prospective isolation and characterization of committed and multipotent progenitors from immortalized mouse mammary epithelial cells with morphogenic potential.

CK5 is uniquely expressed by the clones that possess outgrowth potential. The intensity of CK5 expression was assessed by FACS using a CK5-specific antibody conjugated to Alexa Fluor 488. The histogram shows the intensity of CK5 expression in clones devoid of outgrowth (red) (clones 1, 8, 10, 13 and 14) compared to the clones that formed outgrowths in vivo (blue) (alveolar, ductal and multipotent progenitors). The isotype control is shown in black.

Frances S Kittrell, et al. Breast Cancer Res. 2011;13(2):R41-R41.
4.
Figure 1

Figure 1. From: Prospective isolation and characterization of committed and multipotent progenitors from immortalized mouse mammary epithelial cells with morphogenic potential.

Whole-mount and hematoxylin and eosin staining of outgrowths generated from the CDβ parental line and CDβ progenitor clones. (A and E) Transplantation of COMMA-D cell line engineered to express β-galactosidase (CDβ) parental cell line generates outgrowths containing ductal and alveolar structures. (B and F) Ductal progenitor cells give rise to outgrowths containing ducts only. (C and G) Alveolar progenitor clone generates mainly alveoli and a limited number of small ducts. (D and H) Multipotent progenitor clone generates outgrowths containing ductal and alveolar structures. Figures 1A to 1D are whole-mount-stained images taken at ×3.2 original magnification. Figures 1E to 1H are hematoxylin and eosin-stained images taken at ×10 original magnification.

Frances S Kittrell, et al. Breast Cancer Res. 2011;13(2):R41-R41.
5.
Figure 3

Figure 3. From: Prospective isolation and characterization of committed and multipotent progenitors from immortalized mouse mammary epithelial cells with morphogenic potential.

Immunofluorescence (IF) staining of CDβ-derived progenitor clones showing appropriate basal or luminal orientation. (A to C) CDβ multipotent outgrowths. (D to F) Ductal-limited outgrowths. (G to I) Alveolar-limited outgrowths. (J to L) Adult primary mammary outgrowths (primary BALB/C). (A, D, G and J) Na+, K+, 2Cl- type I cotransporter (NKCC1) (red) and mouse smooth muscle actin (SMA) (green). (B, E, H and K) Cytokeratin 5 (CK5) (red) and CK8 (green). (C, F, I and L) p63 (green) and nuclear estrogen receptor (ER) (red). The nuclei were counterstained with 4', 6-diamidino-2-phenylindole and TO-PRO-3 iodide.

Frances S Kittrell, et al. Breast Cancer Res. 2011;13(2):R41-R41.
6.
Figure 5

Figure 5. From: Prospective isolation and characterization of committed and multipotent progenitors from immortalized mouse mammary epithelial cells with morphogenic potential.

Luminal-restricted clone incorporates into the ductal lumen in vivo and expresses significantly higher luminal-specific markers CK8 and Elf5. (A) The luminal-restricted clone incorporated into the ductal lumen when mixed with primary mammary epithelial cells prior to transplantation. The luminal-restricted cells stained blue upon X-gal staining, since they were derived from CD cells previously engineered to express β-gal. (B) Expression of Elf5 by quantitative real-time polymerase chain reaction is significantly higher in the luminal-restricted clone compared to the remaining progenitor clones. GAPDH, glyceraldehyde 3-phosphate dehydrogenase. (C) Higher percentage of cells in the luminal-restricted clone expressed CK8 compared to the other progenitor clones and nonprogenitor clones 7, 8, 10, and 14. CK8 expression was measured by IF using specific antibody. An average of 500 cells were counted in each group. The x-axis represents the percentage of CK8-positive cells divided by the total number of cells counted. On the y-axis, MP/clone 11 refers to multipotent clone 11 and MP/clone 15 refers to multipotent clone 15.

Frances S Kittrell, et al. Breast Cancer Res. 2011;13(2):R41-R41.

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