HeLa Cell Line Culture Drug Testing: which concentration of three unknown drugs is most effective
by Rhia Buchan
While at Bath University under the supervision of Dr Momna Hejmadi we (Charlotte, Chloe, Laura, Pedro, Will and myself) carry out an investigation on a HeLa cancer cell line (taken originally from Henrietta Lacks who was an African-American women who died on 4th October 1951 from cervial cancer. Her cells from her tumour were then without permission cultured by George Otto Gey and is the first know human ‘immortal’ cell line to be used for medical research).
This investigation entailed growing a three separate cultures of the HeLa cell line in medium. We placed the cells into incubation over night and when we checked them the next day we found that they were growing quicker than expected. Due to this we moved our experiment forward.
To carry out it out we needed a 98 well plate and used it as shown below:
column 1-4 group 1 – Drug A
column 5-8 group 2 – Drug B
column 9-12 group 3 – Drug C
Row G and H were our positive (used bleach) and negative control (had no cells)
The last column in each section was left as a control for each cell culture. We then had to work out the dilution for drug B from the stock (shown below):
Drug B stock 90% – 45×1/90 = 0.5 + 0.5 medium
15×1/90 = 0.16 + 0.84 medium
5×1/90 = 0.05 + 0.95 medium
After working out the dilutions we then diluted the drugs by using medium and p1000 and p20 pipettes as shown below:
- 500 microliters of drug + 500 microliters of medium
- 160 microliters of drug + 840 microliters of medium
- 50 microliters of drug + 950 microliters of medium
We then placed the cells back in incubation at 2:40pm. The next day 11:20am we took the plate out of the incubator. Then on blue towel carefully we blotted the plate to remove the medium then added 100 microliters of MTT then remove the same way the medium was. Then add 100 microliters of isopropanol. This results in the MTT causing crystals to form in intact mitochondria which when isopropanol is added will give a blue colour. Then we looked at the cells under the microscope and analysed the data we obtained.
We found that drug B (Hydrogen Peroxide) was the most effective however would not be an effective cancer treatment drug as it isn’t exclusive in killing only cancer cells but all cells. The next best drug was A (5-Flurorouracil) which is currently used as a cancer treatment and then C (Camptothecin) which is an unproven natural treatment for cancer.