1. The EFFECTS OF OILS ON MG63 CANCER CELLS
Carolyn Guzikowski
Oakland Catholic High School ‘20

2. An Overview of Cancer Cells
Cells that grow and divide at an irregular and unregulated pace
Apoptosis (cells programmed death) stops in cancerous cells; their mutations are passed on to the second generation, and eventually cluster and form tumors
Tumors can either be malignant (aggressive) or benign
Photo Credits: sites.duke.edu

3. MG63 Cell Line Osteosarcoma cells, an aggressive form of bone cancer
Useful model to test the effects of variables on cancer cell proliferation
Photo Credits: neutron.ujf.cas.cz

4. 2 Variables: Essential Oils
Turmeric
Antibiotic
Cancer preventative
Anti-tumor
Antioxidant
Aids in fighting colon cancer
Ginger
Supports healthy digestion
Treats nausea
Helps in occasional joint or muscle discomfort
Photo Credits: up-nature.com and shape.com

5. Purpose
To examine the effects of Ginger Oil and Turmeric Oil on MG63 Cell count proliferation and survivorship.

6. Hypothesis Null Hypothesis Alternate Hypothesis
The different concentrations of Ginger Oil and Turmeric Oil will not affect the cell count proliferation and survivorship.
The different concentrations of Ginger Oil and Turmeric Oil will affect the cell count proliferation and survivorship.

7. Materials
Cryotank
DMEM Media (4 mM L- glutamine, 4500 mg/L glucose, 1 mM sodium pyruvate, and 1500 mg/L sodium bicarbonate + [ 10% fetal bovine serum for complete])
One 75mm2 tissue culture treated flasks
Hemocytometer
Permanent marker
Incubator
Six 25 mm2 tissue culture treated flasks
24 well plate
Aspirating Vacuum Line
Test tube rack
10% fetal bovine serum
Nikon Inverted Compound Optical Scope
Sterile Filter
MG63 Osteosarcoma Cancer Cell Line
Laminar Flow Hood
Trypsin
Laminar Flow Hood UV Sterilizing Lamp
Dye reagent concentrate
Macropipette + sterile macropipette Tips (1 mL, 5 mL, 10, mL, 20 mL)
Spectrophotometer
Labeling Tape
100% Pure Turmeric Oil
Sterile PBS
Micropipettes + sterile tips
100% Pure Ginger Oil
Ethanol (70%)
Distilled water
Nikon Inverted Microscope

8. Combined Variable O (Control) L (Low) H (High) OO OL OH LO LL LH HO HL
Back = Ginger
C = 0 mL
Red = Turmeric
L = .005 mL (5 µl)
H = .05 mL (50 µl)

9. Procedure: Cell Culturing
A 1 mL aliquot of MG63 cells from a Cryotank was used to inoculate 30 mL of 10% serum DMEM media in a 75cm2 culture flask yielding a cell density of approximately 106 to 2x106 cells
The media was replaced with 15 mL of fresh media to remove cryo- freezing fluid and incubated (37° C, 5% CO2) for 2 days until a cell density of approximately 4x106 to 5x106 cells/mL was reached
The culture was passed into 18 flasks in preparation for experiment and incubated for 2 days at 37° C, 5% CO2

10. Procedure: Proliferation Experiment Addition of Variable
Cells from a T75 flask were resuspended after trypsinization to a density of approximately K/mL
4 mL of 10% DMEM media was added to each T25 flask
0.5 mL of cell suspension was transferred to 12 T25 flasks. Flasks were placed back into incubator and cells were allowed to attach for several hours
0.1 mL of Ginger oil was added to 9.9 mL distilled water to create a stock of 1%
Serial dilution to create a 0.01% (stock B) was then created with .1 mL of stock A into 9.9 mL of 10% DMEM. All were sterile filtered using a micron filter
Stock solution process repeated for Low and High Turmeric concentrations
T25 flasks were removed from incubator and the variable was added to reach desired concentrations

11. Variable Concentrations
Proliferation Experiment
Con
Con/Low
Low/Con
Low/Low
Con/High
Cells
0.5 mL
Media
4.5 mL
4.495 mL
4.4 mL
4.45 mL
Stock
0 mL
.005 mL
.01 mL
.05 mL
Total
5 mL

12. Variable Concentrations Cont’d
High/Con
High/High
Low/High
High/Low
Cells
0.5 mL
Media
4.45 mL
4.4 mL
4.445 mL
Stock
.05 mL
.1 mL
.055 mL
Total
5 mL

13. Procedure: Cell Counting
Day 1 and Day 2
The cells were trypsinized and collected into cell 2ml suspension.
10 µl aliquots were transferred to a Hemocytometer for quantification (eight total counts).

14. Statistical analysis of the proliferation results
ANOVA (Single/Double Factor)
Compares variation within groups to variation between groups
Using the ANOVA, a P-value less than the alpha of .05 was gathered (significant variation)
Allows the rejection of the null hypothesis

15. P-Value: 4.11E-16
Ginger

16. P-Value: 3.65E-20
Control
Turmeric
Ginger

17. P-Value: 6.26E-35
Control
Ginger and Turmeric Lows

18. P-Value: 4.55E-35
Control
Low Ginger and High Turmeric

19. Day 1 Counts: Control, Low, High

20. Day 1 Counts: Control Low High

22. Day 2 Counts:

23. Day 2 Counts:

24. D2 LH

25. Conclusions
The null is rejected, Turmeric and Ginger appeared to have significant effects on MG63 Cell count proliferation and survivorship.

26. Future Changes Limitations Extensions Cell counts can vary
Only one cell line used
Use a wider range of concentrations
Compare Turmeric and Ginger to Nigella Sativa (Black Cumin)

27. Works Cited Mark Krotec, PTEI
Zheng, J., Zhou, Y., Li, Y., Xu, D.-P., Li, S., & Li, H.-B. (2016). Spices for Prevention and Treatment of Cancers. Nutrients, 8(8),
MG-63 (ATCC® CRL-1427™). (n.d.). Retrieved February 01, 2018, from aspx#generalinformation