Decellularizing Mammary Fat Pads: Deriving Extracellular Matrix Hydrogels from Mammary Fat Pads
Published: April 30, 2023
Abstract
Source: Alves et. al., Studying Normal Tissue Radiation Effects using Extracellular Matrix Hydrogels. J. Vis. Exp. (2019).
This video describes a technique of deriving extracellular matrix hydrogels by decellularizing murine mammary fat pads following ex vivo irradiation for in vitro studies. This hydrogel helps to mimic the in vivo breast tissue environment post-radiation treatment to study tumor cell behavior.
Protocol
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. On day 1, remove frozen Mammary Fat pads or MFPs from -80 °C and thaw at room temperature. Once thawed, dry MFPs briefly on a delicate task wipe. Weigh the MFPs using an analytical …
Disclosures
The authors have nothing to disclose.
Materials
| 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 |
Cite This Article
Decellularizing Mammary Fat Pads: Deriving Extracellular Matrix Hydrogels from Mammary Fat Pads. J. Vis. Exp. (Pending Publication), e20229, doi: (2023).