Session Title
STEM and Biomedical Research
College
College of Arts and Sciences
Department
Biology
Abstract
Adipose-derived stem cells (ADSCs) are a population of mesenchymal stem cells with multipotent differentiation ability. Three-dimensional cell culture environments are particularly valuable for observing stem cell behavior as they represent a more accurate model of in vivo conditions than simple two-dimensional culture. The purpose of this research was to explore the potential to use several different three-dimensional culture systems to better understand how ADSCs cultured in three-dimensions behave relative to ADSCs cultured using traditional two-dimensional cell culture. The present study hypothesized that both ADSCs both on top of and inside of collagen hydrogels and as spheroids could be cultured to better understand how the cells behave in those environments. It was also hypothesized that cell sorting could be used to obtain a rare subpopulation of ADSCs known as Muse cells. Muse cells have greater developmental potency and the ability to differentiate at a faster rate than non-Muse mesenchymal stem cells. After both two-dimensional and three-dimensional cell culture, real-time RT-PCR was used to analyze gene expression of the housekeeping gene Mrp19 and genes of interest Cxcl12 and Cxcr4. It was observed that ADSCs are amenable to spheroid culture and self-organize into toroids when cultured on top of collagen hydrogels. Several differences were also observed in Cxcl12 and Cxcr4 gene expression between two-dimensional and three-dimensional cultures and between different three-dimensional culture platforms. In addition, the present study was able to identify Muse cells within the ADSC population, which will allow comparison between the behavior of Muse and non-Muse ADSCs in three-dimensional culture in future experiments.
Honors Thesis Committee
Matthew Stern, Ph.D.; Eric Birgbauer, Ph.D.; and LaShardai Brown, Ph.D.
Previously Presented/Performed?
South Carolina INBRE Science Symposium, Columbia South Carolina, January 2020; Sixth Annual Showcase of Undergraduate Research and Creative Endeavors (SOURCE), Winthrop University, April 2020
Grant Support?
Supported by a grant from the South Carolina EPSCoR/IDeA Developmental Research Program, an SC INBRE grant from the National Institute for General Medical Sciences (NIH-NIGMS), and a Ronald E. McNair Post-Baccalaureate Achievement Program grant from the U.S. Department of Education
Start Date
24-4-2020 12:00 AM
Adipose Derived Stem Cell Morphology and Gene Expression in Two-Dimensional versus Three-Dimensional Environments
Adipose-derived stem cells (ADSCs) are a population of mesenchymal stem cells with multipotent differentiation ability. Three-dimensional cell culture environments are particularly valuable for observing stem cell behavior as they represent a more accurate model of in vivo conditions than simple two-dimensional culture. The purpose of this research was to explore the potential to use several different three-dimensional culture systems to better understand how ADSCs cultured in three-dimensions behave relative to ADSCs cultured using traditional two-dimensional cell culture. The present study hypothesized that both ADSCs both on top of and inside of collagen hydrogels and as spheroids could be cultured to better understand how the cells behave in those environments. It was also hypothesized that cell sorting could be used to obtain a rare subpopulation of ADSCs known as Muse cells. Muse cells have greater developmental potency and the ability to differentiate at a faster rate than non-Muse mesenchymal stem cells. After both two-dimensional and three-dimensional cell culture, real-time RT-PCR was used to analyze gene expression of the housekeeping gene Mrp19 and genes of interest Cxcl12 and Cxcr4. It was observed that ADSCs are amenable to spheroid culture and self-organize into toroids when cultured on top of collagen hydrogels. Several differences were also observed in Cxcl12 and Cxcr4 gene expression between two-dimensional and three-dimensional cultures and between different three-dimensional culture platforms. In addition, the present study was able to identify Muse cells within the ADSC population, which will allow comparison between the behavior of Muse and non-Muse ADSCs in three-dimensional culture in future experiments.
