For the first time after 3 years, the SIVB’s In Vitro Animal Cell Sciences Section (IVACS) held an in-person Student and Post-Doctoral Oral Presentation Competition on Monday, June 6, 2022. The top 3 abstracts were chosen for the competition based on their scientific merit and the quality of their write-up. Each presentation was very engaging and was followed by questions from the audience and the competition judges. Our three final contestants delivered professional presentations at a high scientific level. 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. Anissa Belfetmi (Harvard Medical School), Dr. Zoe Zhu (Tufts University), as well as the session moderators Dr. Addy Alt-Holland (Tufts University) and Dr. Kolla Kristjansdottir (Midwestern University). Larissa Marina Pereira Silva, from North Carolina State University/Plants for Human Health Institute, and University of Rio Grande do Norte (UFRN), won the 1st place award for her presentation “A flavonoid-rich extract of moringa oleifera (Moringaceae) leaf cultivated in Brazil inhibited inflammatory mediators in lipopolysaccharide-treated macrophages.” Sepideh Mohammadhosseinpour, from Arkansas State University, won the 2nd place award for her presentation “Assessing the apoptosis effect of prenylated stilbenoids combined with paclitaxel in triple-negative breast cancer cells.” Megan Conkling from Florida Atlantic University – Harbor Branch Oceanographic Institute, won the 3rd place award for her presentation “Habitat Restoration and Production of Bioactive Compounds from 3D Sponge Cell Cultures.” Certificates and monetary awards were presented to the three contestants at the IVACS Business Meeting. Congratulations to these three contestants for their fantastic and engaging presentations!

Submitted by Addy Alt-Holland and Kolla Kristjansdottir

First Place

A Flavonoid-rich Extract of Moringa oleifera (Moringaceae) Leaf Cultivated in Brazil Inhibited Inflammatory Mediators in Lipopolysaccharide-treated Macrophages

Larissa Marina Pereira Silva

Moringa oleifera is a globally recognized herbal medicine due to its pharmacological and nutritional properties, which correlate to the high contents of flavonoids in their leaves. Various persistence diseases are closely associated with excessive inflammation and/or consistent presence of radical oxygen species. Exposure of mammalian cells to LPS leads to the release of pro-inflammatory cytokines and, in turn, activates inflammatory cascades via the secretion of cytokines, lipid mediators, and adhesion molecules, that are common genetic biomarkers involved in the LPS-stimulated murine RAW 264.7 macrophage model inflammatory response in vitro. This present study aimed to evaluate the anti-inflammatory activities of an optimized flavonoid-rich hydroethanolic extract of M. oleifera leaf (MoHLE) and determine the influence of soil cultivation conditions (named T0, T1, T2 and T3) and harvest season of M. oleifera leaf from Northeast of Brazil, in provided anti-inflammatory activities on reactive oxygen species (ROS) and nitric oxide (NO), and inflammatory cytokines in LPS-treated RAW 267.4 cells. As a result the major compounds were characterized by HPLC-ESI-QTRAP-MS/MS in as glycosidic forms derived from apigenin, quercetin, and kaempferol. MoHLE exhibited safety and suppression of generation of nitric oxide (NO) and reactive oxygen species (ROS) between 5 and 50 µg/mL. And down-regulated the transcriptional levels of inflammatory regulatory genes including IL-4, IL-6, IL-1β, COX-2 and iNOS. These findings results showed the potential of this extract as a anti-inflammatory agent to develop new pharmaceutical products, and will contribute to the quality control and standardization of raw materials from M. oleifera, and As far as we know, this is the first attempt to grow open field cultivation of M. oleifera leaves in Brazil. Larissa Marina Pereira Silva, Plants for Human Health Institute (NCRC) and Department of Animal Science, NCSU, Raleigh, NC; and Federal University of Rio Grande do Norte, Natal, BRAZIL. In Vitro Cellular and Developmental Biology, 58:S20, 2022

Second Place

Turning Up the Heat on Editing of the Highly Polyploid Sugarcane Genome

Sepideh Mohammadhosseinpou

Breast cancer is one of the most prevalent types of cancer in women worldwide. Triple-negative breast cancer (TNBC) is unresponsive to typical hormonal treatments causing it to be one of the deadliest forms of breast cancer. Investigating alternative therapies to increase survival rates for this disease is essential. This study aimed to examine if prenylated stilbenoids from peanut can act as an adjuvant for paclitaxel, a chemotherapeutic drug with severe side effects. The prenylated stilbenoids arachidin-1 (A-1) and arachidin-3 (A-3) are analogs of resveratrol (RES) and were produced in hairy root cultures of peanut. The cytotoxicity activity of A-1, A-3, and RES was studied in TNBC cell lines MDA-MB-231 and MDA-MB-436. Furthermore, the cytotoxicity of A-1, the most potent prenylated stilbenoid, combined with paclitaxel was studied by checkerboard assays in the TNBC cell lines. The apoptotic effects of this combination treatment were studied by western blotting targeting protein expression levels of PARP, caspase-8, caspase-9, and survivin and through the Apo-ONE Homogeneous Caspase-3/7 assay. To further investigate the apoptosis and cell cycle stages, cells treated with prenylated stilbenoids or RES were studied using flow cytometry. After 24 hours of treatment, A-1 exhibited higher cytotoxicity than A-3 and RES with approximately 11-fold and 6-fold lower IC50, respectively, in MDA-MB-231 cells, and 9-fold and 8-fold lower IC50, respectively, in MDA-MB-436 cells. A-1 did not show significant cytotoxicity in the non-cancerous cell line MCF-10A. Cytotoxicity, checkerboard, and flow cytometry assays showed a decrease in paclitaxel concentration when combined with prenylated stilbenoids. This highlights the significance of continuing research with prenylated stilbenoids as an adjuvant in TNBC treatment. Sepideh Mohammadhosseinpour, Arkansas State University, PO Box 639, State University, Jonesboro, AR. In Vitro Cellular and Developmental Biology, 58:S19 2022

Third Place

Habitat Restoration and Production of Bioactive Compounds from 3D Sponge Cell Cultures

Megan Conkling

Sponges are ecologically, commercially, and biomedically important organisms, but wild harvest is not enough to meet the demand for research and development of sponge-derived bioproducts. A recent study demonstrated our ability to culture marine sponge cells in three-dimensions (3D) using FibraCel® disks, Ultra-Low Temperature Agarose (ULTA) thin hydrogel layers, and ULTA gel microdroplets (Urban-Gedamke et al. 2021). The creation of a 3D sponge from cryopreserved cells has potential applications for habitat restoration and the production of bioactive materials. Here we report the 3D cultures of three sponges that produce bioactive compounds (discodermolide from Discodermia spp., topsentins from Spongosorites spp., and stevensine from Axinella corrugata). We also report the 3D culture of habitat-building sponges (Xestospongia muta and Geodia cf. gibberosa) found on reefs and in seagrass environments. Cells were successfully cultured in an optimized nutrient medium and formed aggregates that attached to FibraCel® disk fibers as well as within ULTA hydrogel layers. We will discuss the advantages and disadvantages of each method, the production of bioactive compounds from these 3D cultures, and whether these 3D culture methods are a sustainable technique for in vitro production of marine natural products and habitat restoration. Ongoing research is focused on the scale-up of these 3D culture methods to produce marine sponge-derived natural products from other species and to establish nurseries for sponge restoration in habitats impacted by climate change, extreme weather events, or harmful algal blooms. Megan Conkling, Harbor Branch Oceanographic Institute, Florida Atlantic University, 5600 US 1 North, Fort Pierce, FL. In Vitro Cellular and Developmental Biology, 58:S19 2022

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