Lab 2: Trypsinization and Microphotography of Adherent Cells

procedure

handwritten notes

results

<-- HEK 293 cells selected for trypsinization

The plate of cells selected for trypsinization were estimated to be 70%- 75% confluent on June 17, 2023.

<-- HEK 293 cells post trypsinization

4mL of Trypsin/EDTA was added to the plate causing the cells to detach from the plate. The plate was then placed in the incubator for 7 minutes at 37 degrees Celsius. The image was taken on June 17, 2023 after incubation.

<-- 2ml plate of hek 293 cells [4 days post incubation]

2mL of HEK293 cells were added to a new plate containing 10mL of MEM and incubated for 4 days. The picture was taken on June 21, 2023 after being pulled out from the incubator. The cells were estimated to be about 80%-85% confluent and were selected for splitting.

<-- 1.37ml plate of hek 293 cells [4 days post incubation]

1.37mL of HEK 293 cells were added to a new plate containing 10mL of MEM and incubated for 4 days. The picture was taken on June 21, 2023 after being pulled out from the incubator. The cells were estimated to be about 5%-10% confluent indicating that an error had taken place.

<-- 2ml hek 293 cells post trypsinization

4mL of Trypsin/EDTA was added to the plate causing the cells to detach from the plate. The plate was then placed in the incubator for 7 minutes at 37 degrees Celsius. The Image was taken on June 21, 2023 after incubation. The confluency of the cells appears to be greater compared to the first round of trypsinization.

Conclusion

The experiment was a partial success, the 2mL plate of HEK 293 cells exhibited normal growth while the 1.37mL plate contained an error. The error most likely came from a mistake when counting the cells on the hemocytometer or the 1.37mL drawn up was very diluted and contained a small percentage of the cells. To fix this in the future, extra precautions will be taken when counting cells on the hemocytometer and when resuspending the cells to ensure that the cells are evenly distributed throughout the mixture. Otherwise, the 2mL plate of cells had proper growth proving that the growth media made from Lab 1 was also a success.

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