On Monday, June 17, SIVB’s In Vitro Animal Cell Sciences Section (IVACS) held a Student and Post- Doctoral Oral Presentation Competition. The top contestants were chosen to present their research work based on the quality of their abstracts and accomplishments. All the presentations were very professional, and the contestants each fielded questions from both the audience and the judges. While it was difficult to render their decision, the judges, including Dr. Adam Sowalsky, Dr. James Baleja, Mr. Jeffrey Kwak, Ms. Shruti Pore and myself, chose Robert Redden from The Children’s Hospital of Philadelphia as the 1st place winner and Aaron W. Bender from University of Waterloo as the 2nd place winner. Nguyen T.K. Vo from University of Waterloo received an honorable mention certificate for his 3rd place in this competition. Certificates and cash awards were presented at the IVACS Business Meeting. We highly encourage all qualified students and Post-doctoral candidates to submit their abstracts and enroll in the 2014 competition
Submitted by Addy Alt-Holland
First Place
Effects of Differential Trk Expression on Neuroblastoma Cells Cultured in the Microgravity Rotary Bioreactor
Robert A. Redden
Background. Neuroblastoma features confounding biological and genetic heterogeneity that drives diverse, often unpredictable, clinical behavior. Differential expression of the Trk family of neurotrophin receptors strongly correlates with clinical behavior: TrkA expression is associated with favorable outcome, and TrkB with unfavorable outcome. Objective. Neuroblastoma cells cultured in a microgravity rotary bioreactor spontaneously aggregate into tumor-like structures, called organoids. We evaluated the effect of TrkA or TrkB expression on aggregation kinetics and organoid morphology in neuroblastoma cell lines. Methods. SY5Y cells (Trk-null) were stably transfected to express either TrkA or TrkB. Short-term aggregation kinetics were determined by counting the number of single (non-aggregated) viable cells in the supernatant every hour. These data were plotted, and the area under the curve calculated. Organoids were harvested after 4 or 8 days of bioreactor culture, stained, and analyzed morphometrically. Results. SY5Y and SY5Y-TrkB aggregated significantly faster than SY5Y-TrkA cells. SY5Y and TrkB cell lines formed irregularly shaped organoids, featuring stellate projections. In contrast, TrkA cells formed smooth (non-stellate) organoids. SY5Y organoids were slightly smaller on average, but had significantly larger average perimeter than TrkA or TrkB. Conclusion. TrkA expression alone is sufficient to dramatically alter the behavior of neuroblastoma cells in vitro. This pattern is consistent with both clinical outcome and in vivo tumorigenicity, in that SY5Y-TrkA represents a more differentiated phenotype. The microgravity bioreactor is a useful in vitro tool to rapidly investigate the biological characteristics of neuroblastoma and, potentially, as a prognostic assay.
Robert A. Redden, Department of General and Thoracic Surgery , The Children’s Hospital of Philadelphia, Philadelphia, PA. In Vitro Cellular and Developmental Biology, 49:S30, 2013
Second Place
Development of Cell Lines from the Walleye Caudal Fin and Their Ability to Survive and Support Viral Hemorrhagic Septicemia Virus (VHSV) Group IVb Replication at Low Temperatures
Aaron Bender
A unique Northern American genotype (IVb) of viral hemorrhagic septicemia virus (VHSV) emerged over the last 10 years in the Great Lakes, causing significant mortality in a wide range of fish, including walleye. However, cell lines from commercially important species native to this region that could be used to study the virus in these species are few. Therefore, we sought to develop cell lines from the walleye caudal fin and have developed one (WECF11-f) with a fibroblast-like morphology and another (WECF11-e) with an epithelial-like morphology. These survive at temperatures from 4 to 32 °C and provide an opportunity to compare VHSV IVb replication at 4 °C, which walleye experience through winter, with 14 °C, which is usually considered optimal for VHSV production. At 14 °C, VHSV IVb was produced in both cell lines and caused cell death. At 4 °C, the expression of VHSV IVb transcripts was slightly delayed in both cell lines. However, viral production and cell death were greatly delayed at 4 °C but after 2 weeks had occurred in WECF11-f cultures but still had not happened in WECF11-e cultures. Possibly VHSV IVb has a tissue tropism that is most prominently expressed at low temperatures.
Aaron Bender, Department of Biology, University of Waterloo, Waterloo, ON, CANADA. In Vitro Cellular and Developmental Biology, 49:S31-32, 2013
