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Generation of a new bioluminescent model for visualisation of mammary tumour development in transgenic mice

Agnieszka M Zagozdzon1, Patrick O’Leary1, John J Callanan2, John Crown3, William M Gallagher1* and Radoslaw Zagozdzon1

Author Affiliations

1 Cancer Biology and Therapeutics Group, UCD School of Biomolecular and Biomedical Science, UCD Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland

2 UCD School of Veterinary Medicine & Conway Institute, Belfield, Dublin 4, Ireland

3 Molecular Therapeutics for Cancer Ireland (MTCI), c/o National Institute for Cellular Biotechnology, Dublin City University, Glasnevin, Dublin 9, Ireland

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BMC Cancer 2012, 12:209  doi:10.1186/1471-2407-12-209

Published: 30 May 2012

Additional files

Additional file 1:

Figure S1. Identification of potential founders from F0 pups born after microinjections of MMTV-Luc2 construct. Ten mice (blue arrows) were identified by PCR as positive for transgene presence, and five females (green arrows) were selected as potential founders of the MMTV-Luc2 sub-strains #1-5. DNA marker used: 2-Log DNA Ladder (0.1–10.0 kb) (NEB). Figure S2. Whole-body luminescent imaging (upper panels) and ex vivo imaging of isolated internal organs (lower panels) of three representative MMTV-Luc2het virgin females. B: brain, H: heart, Int: intestine, K: kidney, Lg: lungs, Lv: liver, S: spleen, Th: thymus. Figure S3. Distribution of luminescent signal from whole-body imaging of homozygous MMTV-Luc2 littermates. (A) virgin females; (B) males. Mice were of 101 days of age at the time of imaging. Figure S4. Presentation of a representative homozygous MMTV-Luc2 virgin female (A) Distribution of luminescent signal in a whole-body imaging; (B) ex-vivo whole body necropsy; (C) ex vivo imaging of isolated internal organs; B: brain, H: heart, Int: intestine, K: kidney, Lg: lungs, Lv: liver, S: spleen, Th: thymus; (D) representative histological image of the mammary gland of MMTV-Luc2 virgin female. Haematoxylin and eosin stained section of mammary tissue revealing randomly dispersed solitary variable-sized ductular structures embedded in adipose tissue (Bar= 100 μm). (E) and (F) Tomographic surface reconstitution of the luminescent signal from a representative homozygous MMTV-Luc2 female from ventral and dorsal views, respectively. Figure S5. Imaging of the intestine from a representative homozygous MMTV-Luc2 female mouse. The intestine was isolated without prior injection of the luciferin solution to the mouse, 3 therefore the left panel represents spontaneous luminescent signal from the intestine. Then, luciferin solution (300 μg/ml) was applied in drops on the isolated organ and imaging was repeated (right panel). Figure S6. Localisation of the luminescent signal in the whole-body luminescent imaging of F1 females from crosses between homozygous MMTV-Luc2 females and MMTV-PyVThet males. (A) MMTV-Luc2het females (B) Double transgenic and double heterozygous MMTV-Luc2PyVT females. [Note: in panels (A) and (B) mice were imaged for one second and the automatic imaging colour scale adjustments provided by the IVIS Spectrum system were used. The colour bars were removed for the sake of clarity of the figure.] (C) Image obtained with mouse number 27 after setting the exposure time to 20 sec. Figure S7. (A) Whole-body (upper panels) and ex vivo luminescent imaging of mammary tumours (lower panels) isolated from two representative MMTV-Luc2PyVT females at the age of 10 weeks. (B) Histopathological assessment showing evidence of mammary adenocarcinoma formation in a representative MMTV-Luc2PyVT female at the age of 10 weeks. Haematoxylin and eosin stained sections of mammary tumour revealing a multinodular densely cellular mass with cells arranged in sheets and acinar patterns. Increased magnification reveals moderate cell pleomorphism, prominent mitosis and acinar structure formations. (Left-hand image – Bar=500 μm; right-hand image – Bar=25 μm).

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