Collagen gels with embedded fibroblasts did not contract. |
Wrong number of fibroblasts added to the gels. |
Ensure number of fibroblasts 3.5−4.5 × 105 per gel. |
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Fibroblasts died in the gels because of low pH. |
Neutralize the collagen solution with sterile NaOH before adding fibroblasts. |
Some gels contracted whereas others did not. |
Fibroblasts were poorly mixed in the collagen solution. |
Mix fibroblasts with the collagen solution by gently inverting the tubes. |
Poor epithelial differentiation. |
Density of keratinocytes on the gels was too low. |
Seed proper number, 1.0−1.2 ×106 oral epithelial cells on the gels. |
|
Failure to maintain cultures on air-liquid interface. |
Check cultures daily for the medium levels. Aspirate media from inside the inserts. |
None, few or disturbed epithelial layers. |
Cultures were disturbed while placing into the cassettes or during during histological processing. |
While harvesting the cultures, do not disturb them. Make sure the cultures do not move along the sponges in the cassettes. |