Question

Describe an experiment to compare the stiffness effects of cell vs. extracellular matrix on stem cell using hydrogels.

Answer 1

For regenerative medicine Mesenchymal stem cells are valuable cell source.

For the differentiation of these cells the interaction between the cells and the extracellular matrix play a great role. Cells are very sensitive to the mechanical properties of extracellular matrix such as stiffness. The range of stiffness varies greatly from soft, pliable to very hard. To perform an experiment first Collagen-I gel was prepared after neutralising the collagen-I with the help of NaOH. The soft substrate had collagen-I gel with about 500 μm thickness. Before polymerization the collagen-I gel solution is used for the culture dishes so that they get rinsed and excess solution can be removed which resulting in a thin layer coating of polymerized collagen then polyacrylamide gels with different amount of crosslinker is used. Treatment of glass slides is done with a chemical named 3-aminopropyltrimethoxysilane and 0.5% glutaraldehyde.

The gel allowed to polymerise so that it forms thick gel on slides. Chemical named Sulpho-SANPAH can be used to link collagen-I to polyacrylamide gel. Substrates get sterilised by UV. With the help of Comparator the elastic modulus of polyacrylamide gel can be determined. The comparator can measure the displacement of a gel in response to certain force

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