Talk:20.109(S11):Testing cell viability (Day3)

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T/R schedule

Staining protocol time is expected to be ~ 1 hour, leaving 25 min blocks of microscope time for each group.

Please come prepared to lab (i.e., having read the protocol), and feel free to arrive even earlier than the suggested times below.

Microscope time Arrival time (at latest!) Team colour
2:05-2:30 1:05 Yellow
2:30-2:55 1:20 Red
2:55-3:20 1:45 Blue

mesenchymal stem cells grown in low viscosity alginate with various conditions

0.5mg/mL HA and no collagen added. Image was taken with the bead core up.
0.5mg/mL HA and no collagen added. Image was taken with the bead core up.
0.5mg/mL HA and approx. 0.1% collagen added. Image was taken with the bead core down. Bead was squished by the coverslip, resulting in lower density of cells in the field of view.
0.5mg/mL HA and approx. 0.1% collagen added. Image was taken with the bead core down. Bead was squished by the coverslip, resulting in lower density of cells in the field of view.
3:20-3:45 2:10 Purple

mesenchymal stem cells grown in low viscosity alginate with various conditions

0.05mg/mL HA and no collagen added
0.05mg/mL HA and no collagen added
0.05mg/mL HA and collagen II added
0.05mg/mL HA and collagen II added
3:45-4:10 2:35 Pink

Testing chondrocytes with and without the presence of ascorbate (a growth factor). Beads a made from low viscosity alginate.

Live cells shown with ascorbate added
Live cells shown with ascorbate added
Few cells can be seen in the sample without ascorbate, most likely due to cells falling out of the bead.
Few cells can be seen in the sample without ascorbate, most likely due to cells falling out of the bead.
4:10-4:35 3:00 Orange
4:35-5:00 3:25 Green

W/F schedule

Staining protocol time is expected to be ~ 1 hour, leaving 25 min blocks of microscope time for each group.

Please come prepared to lab (i.e., having read the protocol), and feel free to arrive even earlier than the suggested times below.

Microscope time Arrival time (at latest!) Team colour
2:05-2:30 1:05 Purple We are testing chondrocytes in 2% low viscosity alginate with different CaCl2 concentrations.
Beads prepared in 20mM CaCl2. Picture taken of the bead core.
Beads prepared in 20mM CaCl2. Picture taken of the bead core.
Beads prepared in 200mM CaCl2. Picture taken of the bead core.
Beads prepared in 200mM CaCl2. Picture taken of the bead core.
2:30-2:55 1:20 Pink We tested a cell density of around 1e7 cells in 1.5% low viscosity and 2e6 in 1.5% medium viscosity.
Surface of 1.5% low viscosity bead.
Surface of 1.5% low viscosity bead.
Surface of 1.5% medium viscosity bead.
Surface of 1.5% medium viscosity bead.
2:55-3:20 1:45 Blue We tested the impact of different alginate viscosities on chondrocyte differentiation. Cell density was 1 million cells/mL in 1.5% low and 1.5% medium viscosity alginate.
Surface of 1.5% low viscosity bead.
Surface of 1.5% low viscosity bead.
Center of 1.5% medium viscosity bead.
Center of 1.5% medium viscosity bead.
3:20-3:45 2:10 Green We wanted to see if seeding a stem cell culture with chondrocytes could increase the rate of differentiation. Cell density was 3.6 million cells/mL in 1.5% low viscosity alginate.
Core of chondrocyte-only bead.
Core of chondrocyte-only bead.
Core/Edge of stem-cell only bead. The left, near the edge, has many dead cells, while the right, near the core, has mostly alive cells.
Core/Edge of stem-cell only bead. The left, near the edge, has many dead cells, while the right, near the core, has mostly alive cells.
Core of 2:1 Chondrocyte:stem cell bead.
Core of 2:1 Chondrocyte:stem cell bead.
3:45-4:10 2:35 Orange We were testing the influence of BMP-6 upon the differentiation of mesenchymal stem cells into chondrocytes. Cells were cultured in 1% medium-viscosity alginate.
Center of the control bead.
Center of the control bead.
Center of the BMP-6 exposed bead.
Center of the BMP-6 exposed bead.
4:10-4:35 3:00 Yellow We tested chondrocytes on 1.5% medium-viscosity alginate prepared in regular control conditions, or with addition of 800 uM glycine, a major component of collagen.
Center of the control (no glycine) bead.
Center of the control (no glycine) bead.
Center of the bead under +glycine condition.
Center of the bead under +glycine condition.
4:35-5:00 3:25 Red We tested chondrocytes on 1% medium-viscosity alginate prepared in either 20mM or 200mM CaCl2 by Live/Dead Fluorescence Assay.
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