Fabrication of Extracellular Matrix from Mouse Model: An Effective Procedure to Prepare Decellularized Tongue Extracellular Matrix from Murine Tongue
Transcrição
The extracellular matrix or ECM – the non-cellular component of the tissue environment – consists of a complex network of structural and regulatory proteins and polysaccharides secreted by cells. The ECM provides structural and biochemical support to cells and regulates their growth.
To prepare the tongue extracellular matrix, begin by taking a freshly harvested mouse tongue. Wash it with ethanol to remove its lipid layer.
Next, subject it to alternating freeze and thaw cycles. Freezing promotes intracellular ice crystal formation, while thawing melts the ice crystals, resulting in the disruption of the cellular membrane.
Now, suspend the tongue in an antibiotic solution to inhibit bacterial growth. Subsequently, treat the organ with a hypertonic saline solution. In the presence of a high solute concentration, water flows out from the cells, causing them to shrink. This step aids in cellular death and detachment from the extracellular matrix.
Incubate the tongue in a non-ionic surfactant. The surfactant completely solubilizes the cellular and nuclear membranes, facilitating the release of intracellular contents into the solution.
Lastly, suspend the tongue in a solution containing calcium and magnesium ions, followed by nucleases treatment. The calcium and magnesium ions enhance the activity of nucleases to degrade any free nucleic acids in the solution.
Store the decellularized tongue tissue in a suitable buffer until further processing.
