Talk:20.109(S10):Initiate cell culture (Day2)

Experimental Plans and Predictions

T/R Lab

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Experimental plan: Add .66 mmol sodium bicarbonate per 1 mL media (total 1.8 mL bicarbonate in 6 mL media) to raise the pH of the media to 8.0.

Cell type: chondrocytes

Cell density: 10^6 cells/6.4 mL

Total # cells needed: 6 million

Standard media other than bicarbonate addition

scaffold: Sigma Aldrich low viscosity 2% high M

Expectations: We expect the viability of the chondrocytes to drop at the higher pH; however, we do not expect the pH to be so far out of the biological range as to completely kill the chondrocytes. We expect the cells at biological pH (7.2-7.4) to maintain chondrocyte phenotype better than the cells at high pH.

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[Pink:] Since mechanical loading and pressure in the joints is one of the factors that contributes to cartilage degradation in secondary osteoarthritis, through our experimental design, we seek to evaluate the effect of compression and pressure on the ability to grow a 3D chondrocyte culture. We will grow two cultures in a six well plate: both cultures will be covered with a square glass slide (m = 0.189 g, A=22 mm x 22 mm), and one of them will also be subjected to additional pressure by placement of a metal mass (m= 24.608 g) on top of the glass slide. We think that the control (weightless) sample will preserve a chondrocyte-like phenotype, whereas the compressed sample will lose chondrocyte phenotype and will possible not be as viable, since we think compression and confinement in the scaffold will contribute to chondrocyte degradation. (Cell culture conditions: Sigma Aldrich "low viscosity" alginate at 1.5%; Differentiated chondrocytes, 10^7 cells/mL, media conditions are standard).

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W/F Lab

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