1. Electromagnetic Field Effects on MG-63 Cell Lines
Napat Kulruchakorn
Pittsburgh Central Catholic
Grade 11

2. Electromagnetic Field
Magnetic Field produced by electrically charged objects
Common in today’s environment from wireless waves produced by technological devices

3. Electromagnetic Field in a Solenoid
Uniform throughout the structure
Flows in one direction
Highly concentrated within the coil

4. An Overview of Cancer Cells
Cancer cells are cells that grow and divide at an irregular, unregulated pace.
Apoptosis does not occur in cancerous cells; their mutations are passed on to the second generation, eventually clustering and forming tumors.
Tumors can be malignant (aggressive) or benign.

5. MG-63 Cancer Cell Line Human cancer cell line
Osteosarcoma cells, an aggressive form of bone cancer
Useful model to test the effects of variables on cancer cell proliferation

6. Past Studies
In cancer cells, the membrane potential level is significantly lower than that of healthy cells
Cancer cells are more detached and do not exhibit contact inhibition of their growth
Therefore, could charged particles within the membrane of the cancer cells be influenced by external electromagnetic forces?

7. Purpose
To examine the effect of electromagnetic fields on MG-63 cell proliferation and survivorship

8. Hypotheses
Null: The different strengths of electromagnetic fields will not have a significant effect on the proliferation and survivorship of the MG-63 cells.
Alternative: The different strengths of electromagnetic fields will have a significant effect on the proliferation and survivorship of the MG-63 cells.

9. Materials The Solenoid: Insulated Copper Wire 6V Lithium Battery
Water Bottle (for structure)
Aluminium Foil
Cable Zip Ties
Multimeter

10. Materials (cont’d)
Cryotank
Fetal Bovine Serum (FBS)
MG-63 Cell Line
Trypsin-EDTA
Pen / Strep
75 mm2 tissue culture treated flasks
25 mm2 tissue culture treated flasks
Macropipette + Sterile Macropipette Tips (1 mL, 5 mL, 10 mL, 20 mL)
Micropipette + Sterile Micropipette Tips
DMEM Serum - 1% and Complete Media (4mM L-glutamine, mg/L glucose, 1 mM sodium pyruvate, and 1500 mg/L sodium bicarbonate + [10% fetal bovine serum for complete])
75 mL culture flask
Incubator
Nikon Inverted Microscope with Imaging Technology
Laminar Flow Hood
Laminar Flow Hood UV Sterilizing Lamp
Hemacytometer
Sterile PBS
Ethanol (70%)
Ethanol (70%)
Sharpie Pen
Sterile Water
Purple Nitrile Gloves

11. Procedure (Constructing the Solenoid)
For each solenoid:
A plastic water bottle was used as a structure.
The bottle was wrapped with one layer of Aluminum Foil
A 61-meter insulated copper wire was coiled 296 times around the circumference, creating a cylindrical structure
The wire coil was held together with cable zip ties
The batteries were prepared to meet their exact charges. Each would run out after 10 hours of exposure time.

12. Procedure (Cell Line Culture)
A 1 mL aliquot of MG63 cells from a Cryotank was used to inoculate 30 mL of 10% serum DMEM media in a 75mm2 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 8 flasks in preparation for experiment and incubated for 2 days at 37° C, 5% CO2

13. Procedure (Addition of Variable on Day 0)
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 8 T25 flasks. Flasks were placed back into incubator and cells were allowed to attach for several hours

14. Procedure (Addition of Variable on Day 0)
Each end of the insulated copper wire was connected to the cathodes and anodes of the 6V and 12V batteries under a sterile environment.
T25 flasks and solenoids were incubated at 37C and 5% CO2.

15. Procedure (Cell Counts)
Day 1 and Day 3
Cell densities were determined as follows:
The cells were trypsinized and collected into cell suspension.
20 μl aliquots were transferred to a Hemocytometer for quantification (eight total counts).

16. Strength of Magnetic Field
Field Strength Levels
Turn Density
Strength of Magnetic Field
Current
Permeability (Air)
= 4π × 10-7

17. Field Strength Levels High (12V) Low (6V) Control Tesla Gauss 2.2282
Gauss
2.2282
1.1141
× 10−4
× 10−4

18. Electromagnetic Field Effects on MG-63 Proliferation
P-Value Day 1: 3.8x10-12
P-Value Day 3: 6.1x10-10

19. Dunnett’s Test Day 1 Day 3 Field Strength T Value T Crit Results High
2.88
Significant
Low
Day 1
Field Strength
T Value
T Crit
Results
High
2.88
Significant
Low
6.7817
Day 3

20. Conclusion
The null hypothesis was rejected. The data from Day 1 and Day 3 strongly suggested that both low and high strength levels of electromagnetic field have significant effects on the survivorship and proliferation of MG- 63 cells.

21. Limitations
The batteries needed to be small enough to fit in the incubator, resulting in short exposure time
Only two different strength levels tested
Only one cell line used
Low number of flasks
Cell health not observed
Cell counts can vary

22. Project Improvements
An alternative power source that would allow longer exposure time
More flasks
Use multiple cell lines
More strength levels of EM fields

23. Citations / Acknowledgements
Mark Krotec, PTEI
Bob Leonard
Presman, A. S. Electromagnetic Fields and Life. New York: Plenum, Print.
Steve Haltiwanger M.D., C.C.N. The Electrical Properties of Cancer Cells (2002): Web.
Cone CD. The role of surface electrical transmembrane potential in normal and malignant mitogenesis. Ann NY Acad Sci 1975;238:

24. ANOVA - Day 1 Analysis of Variance (One-Way) Summary Groups
Sample size
Sum
Mean
Variance
Low 8
21,120,000
2,640,000
E+11
High
984,000
123,000
1,000,285, C
44,790,000
5,598,750
E+11
ANOVA
Source of Variation SS df MS F
p-level
F crit
Between Groups
E+14 2
E+13
E-12
3.4668
Within Groups
E+12 21
E+11
Total
E+14 23

25. ANOVA - Day 3 Analysis of Variance (One-Way) Summary Groups
Sample size
Sum
Mean
Variance
Low 8
24,030,000
3,003,750
E+11
High
633,000
79,125
951,267, C
56,490,000
7,061,250
E+12
ANOVA
Source of Variation SS df MS F
p-level
F crit
Between Groups
E+14 2
E+13
E-10
3.4668
Within Groups
E+13 21
E+12
Total
E+14 23

26. Concentrations (cells / mL)
Day 1
Low
High
Control
144000
69000
129000
135000
99000
102000
171000
Day 3
Low
High
Control
105000
129000
30000
90000
51000
81000
66000