John Sibbitt


Education: Bachelor of Science in nutrition science (May 2014)

Currently pursuing a Professional Science Master in applied biosciences at Kansas State University

McNair Project: Determining Effective Culturing Method for PC346-C Cell Line for In-Vitro Study of Prostate Cancer (2011)

Mentor: Brian Lindshield, Ph.D.

Background: Commonly, aggressive and fatal prostate carcinomas evolve from PC's initial mild nature, making it the second leading cause of male cancer deaths in America. Therefore, a cell line resembling early stage PC has importance in clinically relevant in-vitro research. The majority of diagnosed PC's are initially mild in nature, thus understanding PC's progression to more advanced stages can prove pivotal in prevention of deadly carcinomas. The PC346-C cell line is unique, because it maintains early stage PC characteristics, unlike the majority of PC cell lines, which are derived from and model advanced stages of the disease. However, PC cells are among the most difficult to sustain in culture. To help, we documented potential pitfalls of culturing viability and possible solutions.

Approach/Results: PC346-C cells were obtained from the Erasmus Medical Center, located in Rotterdam, Netherlands. Cells were taken up from cryopreservation and seeded into Primiria T-25 flasks. Flasks contained 4 mL of DMEM-F12 base media plus 17 different complex additives to encourage attachment and growth. Flasks were incubated at 37 Celsius in a 5% C02 atmosphere and media were changed every 3 days. If confluence was reached, cells were sub-cultured via trypsinization at 1:3 split ratios. Attachment periods, percentages of confluence, doubling times, and general cell health were documented to ascertain the culture's viability. The cells displayed healthy morphology; however, proliferation rates were sub-optimal.

Conclusions: Human error in the preparation of the complex media likely contributed to the poor growth observed. After identifying several growth inhibiting problems, we boosted proliferation rates. However, proliferation was still sub-optimal. Further study is needed to determine the reason for these poor growth rates. The difficulty of culturing early stage prostate cancer in-vitro models was verified.