2009 SIVB Student Awards

The following student awards were presented at the 2009 In Vitro Biology Meeting, Charleston, South Carolina. Information related to the available specific student awards can be found on the SIVB website (www.sivb.org) or by contacting the SIVB Business Office at (919) 420-7940, sivb@sivb.org, or Dr. Pamela Weathers, Chair, Student Affairs and Awards Committee, at weathers@wpi.edu.

Nicole Napolitano

2009 HONOR B. FELL AWARD, HOPE E. HOPPS AWARD, AND STUDENT TRAVEL AWARD

Collagen Production by Normal Human Dermal Fibroblast Cells in Serum-free Media

Collagen is a protein found in the skin and bones that is associated with supporting cell structure and strength. Increased collagen production has been shown to advocate healthier, younger skin, and aid wound healing. Assays to help discover collagen inducing compounds would be valuable to academia and biotechnology companies. The purpose of this study was to establish an assay that measured collagen production of normal human dermal fibroblasts (NHDF) grown in standard serum media and serum-free supplemented media. Ascorbic acid has been shown to potently induce collagen production in fibroblasts. NHDF were grown in standard serum media in the absence or presence of ascorbic acid (150ug/mL) for 7 days and 14 days. Cells were also grown in chemically enhanced serum-free supplemented media under the same testing conditions. The serum free media minimized interactions with the collagen assay while providing several growth factors to help maintain cell function. Collagen was measured using Sirius Red dye, a collagen specific binding dye. Collagen was measured in the growth media and on the matrix. NHDF grown in the chemically enhanced media and ascorbic acid showed a 2 and 3.5 fold increase of collagen on the matrix at day 7 and 14 respectively, compared to the 1.5 and 2.5 fold increase seen in NHDF grown in standard media. Also at day 14, NHDF grown in the chemically enhanced media with ascorbic acid showed a 2.5 fold increase of collagen in the media. Using this assay, several compounds were tested for the ability to increase collagen production in NHDF grown with chemically enhanced media. Although none of the compounds showed a potent induction of collagen production, there were subtle changes compared to the media control. The Sirius Red Assay successfully measured collagen production in the growth media and on the extracellular matrix. Data suggests that the chemically enhanced media is a good alternative to serum based media and is able to support collagen induction in cells. This model can be used to discover new collagen inducing compounds and expanded to study other skin related proteins.

Nicole Napolitano, Department of Pathology, Stony Brook University, Stony Brook, NY 11749. In Vitro Cellular and Developmental Biology, 45:S49-50, 2009

Sean Michael Halloran

2009 PHILLIP R. WHITE AWARD AND STUDENT TRAVEL AWARD

Macronutrient Optimization for In Vitro Growth of Turmeric (Curcurma longa L.)

Most media for plant tissue culture is based on Murashige and Skoog 1962. This work maximized the fresh weight of tobacco callus on agar (one nutrient at a time) with few second order interactive effects. Our current work combined powerful statistical software designs to analyze the complex interactions of macronutrients in liquid culture micropropagation of turmeric (Curcurma longa L.), an important herbaceous perennial. The reasons for re-evaluating MS included: differences of water availability in agar and liquid media; media have not been defined for commercially micropropagated herbaceous perennials; differentiated tissues have greater nutrient requirements than callus; the limited potential of prior workers to conduct and analyze proper experiments to measure these interactions; modern designs save labor, space, materials, and increase visualization of multi-factor interactions. This experiment is a d-optimal design with 3 levels of sucrose (1.5 - 6%), plants per vessel (3 - 9 plantlets), volume (25 - 45 mL), nitrate (10 - 50 mM), and potassium and nitrogen expressed as a 2-component mixture. The full factorial of all these variables is an impractical experiment, while our d-optimal design space required only 55 vessels. Greatest plant multiplication in vitro was with 10 mM nitrate and 1:1 ammonium/potassium (25% of MS), 3 plants per vessel, and 6% sucrose. At higher nitrogen levels, like MS 1962, the sucrose and plant density are not apparent. This maximization shows the significance of analyzing the second order interactions to optimize factors involved in plant growth. The quality of plantlets to grow in the greenhouse likely has a different optimization. An evaluation of dissimilar combined responses will be made increased sensitivity of chemical exposures will be presented.

Sean Michael Halloran, Department of Horticulture, Clemson University, Clemson, SC 29634. In Vitro Cellular and Developmental Biology, 45:S60, 2009

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