The SIVB’s In Vitro Animal Cell Sciences Section (IVACS) held the virtual Student and Post-Doctoral Oral Presentation Competition on Monday, June 7, 2021. The top 3 abstracts were chosen for the competition based on their scientific merit and the quality of their write-up. Although the competition was virtual, due to COVID-19 pandemic, each presentation was very engaging and at a high scientific level. Our three final contestants delivered professional pre-recorded presentations and continued lively on-line discussions with the audience and judges thought the virtual conference. The panel of IVACS judges that evaluated the presentations included: Dr. John W. Harbell (JHarbell Consulting, LLC), Dr. Barbara Doonan (New York Medical College), Dr. Michael J. Fay (Midwestern University), Dr. Brad L. Upham (Michigan State University), Dr. Michael K. Dame (University of Michigan), Dr. Mae Ciancio (Midwestern University), Dr. Kristina Martinez-Guryn (Midwestern University), and Anissa Belfetmi (Harvard Medical School), as well as the session moderators Dr. Addy Alt-Holland (Tufts University) and Dr. Kolla Kristjansdottir (Midwestern University). Elizabeth Hartley Urban-Gedamke, from Florida Atlantic University – Harbor Branch Oceanographic Institute, won the 1st place award for her presentation “Successful Culture of Marine Sponge Cells Using Multiple 3-D Culture Methods” Renato C. Aguilera, Jr., from Tufts University, won the 2nd place award for his presentation “Multidimensional Tomography of Cells by a Combination of Atomic Force Microscopy and Dynamic Mechanical Spectroscopy. ” Stephanie Lourdes Echeverria, from Midwestern University, won the 3rd place award for her presentation “Assessing CCR1 Antagonists for Chemotaxis Inhibition in a Multiple Myeloma in Vitro Model.” Certificates and monetary awards were sent to the three contestants via mail. Congratulations to these three contestants for their fantastic virtual presentations!

Submitted by the Business Office on behalf of Addy Alt-Holland and Kolla Kristjansdottir

First Place

Successful Culture of Marine Sponge Cells Using Multiple 3-D Culture Methods

Elizabeth Urban-Gedamke

Sponges are ecologically, commercially, and biomedically important organisms, but the requirements for sponge tissue for research, restoration, and pharmaceutical production cannot be met by wild harvest alone. A recent study demonstrated the ability to culture marine sponge cells in two-dimensions using an optimized nutrient medium. Here we report the successful implementation of the same optimized nutrient medium to produce three-dimensional cell cultures of the marine sponge Geodia neptuni using four different methods – FibraCel© discs, thin hydrogel layers, gel micro droplets, and spheroid formation. Herein we discuss the advantages and disadvantages of each method, as well as make recommendations for future applications. The research furthers our ability to study and culture marine sponge cells, and optimization of these techniques may lead to the in vitro production of sponge cultures for the synthesis of marine natural products and for habitat restoration purposes.

Elizabeth Urban-Gedamke, Florida Atlantic University – Harbor Branch Oceanographic Institute, Ft. Pierce, FL. In Vitro Cellular and Developmental Biology, 57:S32, 2021

second Place

Multidimensional Tomography of Cells by a Combination of Atomic Force Microscopy and Dynamic Mechanical Spectroscopy

Renato Aguilera

Cell phenotyping by their mechanical properties is gaining increasing interest as a type of label-free biomarker. Alterations to the cytoskeleton and nucleus due to cell instability, passage, or differentiation lead to significant changes in cell mechanics. Additionally, precise mechanical measurements enable viscoelastic modeling of cell deformation for high-speed rheological studies. Here, we present the application of our new technique, FT-NanoDMA, to create a multidimensional tomography of the dynamical mechanical properties of cells. FT-NanoDMA stands for Fourier transformed nano dynamical mechanical analysis, which is implemented by means of atomic force microscopy (AFM). It allows for simultaneous recording at different frequencies of multiple dynamical mechanical properties of cells, such as the storage and loss moduli, loss tangent, etc.  The FT-nanoDMA is a modality that allows for mechanical analysis of nanointerfaces and single cells with a nanoscale spatial resolution (up to 10-70 nm when recorded on fixed cells) and spans the entire biological spectroscopic range (up to 300 Hz). The images of the distribution of these parameters can be recorded at each particular depth of the cell, thereby creating a stag of images suitable to create a multidimensional tomography of single cells. We demonstrate examples, which show that the tomographic images show unique features not present in the complementary fluorescent/optical images of cells. The biological significance of this novel information has yet to be understood.

Renato Aguilera, Tufts University, Medford, MA. In Vitro Cellular and Developmental Biology, 57:S32-33, 2021

third Place

Assessing CCR1 Antagonists for Chemotaxis Inhibition in a Multiple Myeloma in Vitro Model

Stephanie Lourdes Echeverria

Multiple myeloma (MM) is a plasma B-cell malignancy characterized by osteolytic bone lesions. MM cells secrete and express CCL3/MIP1α which upregulates osteoclastogenesis. Elevated CCL3 levels display a chemotactic ability on isolated osteoclast precursors. Increased levels of CCL3 in MM patients correlates with a greater disease burden, due to the increase of bone resorption, and is indicative of a worse prognosis when compared to MM patients exhibiting lower CCL3 levels. CCR1, a GPCR chemokine receptor, is endogenously expressed on MM cells, and can bind with CCL3. In a previous study, the RPMI8226 MM cell line, CCL3-mediated CCR1 chemotaxis was inhibited in a dose-dependent manner by six CCR1 antagonists (AZD4818, BX471, CCX354, CP481715, MLN3897, PS031291). In this study, we assessed the MM cell line U266 as well as a transfected cell line, U266_CCR1. While U266 cells express lower levels of CCR1 than RPMI8226, U266_CCR1 express higher levels of CCR1.  Cells were treated with a serial dilution of the same six CCR1 antagonists and chemotaxis to either supernatant from osteoclast precursor RAW 264.7 cells or Fetal Bovine Serum (FBS) in which a multitude of growth factors and chemokines are present was examined. We hypothesized the six CCR1 antagonists would result in a dose-dependent inhibition of chemotaxis similar to that seen with RPMI8226. Instead, we found only two of the compounds (AZD4818 and BX471) inhibited chemotaxis of the U266 and U266_CCR1 MM cell lines towards RAW 264.7 supernatant or FBS. For both AZD4818 and BX471 there were differences between the chemotactic response of the U266 and U266_CCR1 cell lines, with the U266_CCR1 cell line having a greater degree of inhibition, suggesting the inhibition is driven by CCR1.

Stephanie Lourdes Echeverria, College of Graduate Studies, Midwestern University, Downers Grove, IL 60515. In Vitro Cellular and Developmental Biology, 57:S32 2021

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