Decellularizing Mammary Fat Pads: Deriving Extracellular Matrix Hydrogels from Mammary Fat Pads

Protokoll

1. Decellularization

NOTE: This procedure was adapted from previously published methods focused on adipose decellularization, which included the sodium deoxycholate ionic detergent rather than sodium dodecyl sulfate to remove DNA efficiently.

1. On day 1, remove frozen Mammary Fat pads or MFPs from -80 °C and thaw at room temperature.
2. Once thawed, dry MFPs briefly on a delicate task wipe. Weigh the MFPs using an analytical scale.
3. Using a pair of forceps with scissors or a scalpel, divide tissue up into 3 mm x 3 mm x 3 mm samples for study of the intact ECM and ....

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Materialien

Name	Company	Catalog Number	Comments
10% Neutral Buffered Formalin, Cube with Spigot	VWR	16004-128
2-methylbutane	Alfa Aesar	19387
Bovine Serum Albumin	Sigma-Aldrich	A1933-25G
Dulbecco's phosphate-buffered saline	Gibco	14040133
Fisher Healthcare Tissue-Plus O.C.T. Compound	Fisher Scientific	23-730-571	cryostat embedding medium
Kimtech Science Kimwipes	Kimberly Clark		delicate task wipes
n-Propanol (Peroxide-Free/ Sequencing), Fisher BioReagents	Fisher Scientific	BP1130-500
PBS (10X), pH 7.4	Quality Biological, Inc.	119-069-151	Phosphate-buffered saline
Penicillin-Streptomycin	Gibco	15140-122
Peracetic acid	Sigma-Aldrich	77240-100ML
Triton x-100	Sigma-Aldrich	X100-100ML	t-Octylphenoxypolyethoxyethanol
Trypsin-EDTA (0.25%), phenol red	Gibco	25200-056
Richard-Allan Scientific Signature Series Hematoxylin 7211	Richard-Allan Scientific	7211
Whatman qualitative filter paper, Grade 4	Whatman	1004-110	grade 4 qualitative filter paper
Xylenes (Certified ACS), Fisher Chemical	Fisher Scientific	X5-5