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

Experimental Plans and Predictions

T/R Lab

Red:

Orange:

Yellow:
| colspan="3" |

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.	chondrocytes	[[]]	[[]]	[[]]

|- style="height:200px" |

|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.

Green:

Blue:

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.

Purple:

W/F Lab

Red:

Orange:

Yellow:

Green:

Blue:

Pink:

Purple: