Lab Report

1146 WordsNov 1, 20175 Pages
which would have only surface-seeded cells. The well plate was wrapped in Parafilm to prevent contamination and floated in a 37 C water bath for 30 min to allow crosslinking. Cell solution was then added to the collagen gels, with the amount again dependent on treatment—83 μL was added to wells with only encapsulated cells or only surface-seeded cells; 41:5 μL was added to wells with both encapsulated and surface-seeded cells. 108:5 μL of collagen solution was added to gels with encapsulated cells, and 41:5 μL of cell solution was then added to the gels with both encapsulated and surface-seeded cells. The gels were then incubated for an additional 30 min before the washers were removed with sterile forceps, completing the gel preparation…show more content…
A two-sided t-test for difference between means was used to assess statistical significance between observed differences in contraction. Results and Discussion Gel Compaction Both encapsulated and surface-seeded cells were observed to contract the collagen lattice and exclude water, causing a compaction of the gel observable at a macroscopic level consistent with previous reports.4 This compaction was not observed in gels without seeded cells. The gels were reasonably stiff and safely handled without much concern. After two days, the color of the gels did not change noticeably from the original near-transparence; however, it is possible that the previously reported color change would have been more pronounced after a longer time interval.4 The rate of collagen contraction was observed to depend upon cell seeding location, with encapsulated cells contracting the surrounding lattice more rapidly than surface-seeded cells (Figure 1), as hypothesized. This makes intuitive sense, since the surface-seeded cells have a sort of induced 3 Figure 1: Graphs summarizing the observed compaction of collagen gels for each treatment. apical-basal polarity, as the gel exists only on the basal side of these cells, so they can contract the lattice only in this direction, whereas encapsulated cells have no induced polarity and can contract the lattice isotropically. However, isotropic contraction was assumed in all cases partly to simplify the mathematics and partly

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