Spheroid Culture of Murine and Human Adipose-Derived Stem Cells Alters the Expression of Genes that Regulate Developmental Potency

Poster Number

046

College

College of Arts and Sciences

Department

Biology

Faculty Mentor

Matthew Stern, Ph.D.

Abstract

Adipose-derived stem cells (ADSCs) are multipotent, mesenchymal stem cells that are found within the microvasculature of adipose tissue. While ADSCs have the potential to differentiate into multiple cell lineages, they cannot match the differentiation potential of pluripotent stem cells, such as embryonic stem cells or induced pluripotent stem cells. Previous research has shown that stem cells are sensitive to the composition and dimensionality of their culture environment. We have shown that culturing murine ADSCs as three-dimensional spheroids alters their gene expression compared to ADSCs cultured using traditional, two-dimensional methodology. We hypothesized that human ADSCs cultured as spheroids and in two dimensions would also exhibit differential gene expression, and that the changes in gene expression would mirror those observed in murine ADSCs. To investigate our hypothesis and optimize our protocol for generating spheroids, we compared our traditional hanging-drop method to the use of micropore plates that are specially designed for producing spheroids. Our results suggest that human ADSC gene expression is significantly altered by spheroid culture, with some changes mirroring what was observed in murine ADSCs. The method of spheroid generation had little impact on the results; however, the workflow associated with use of the micropore plates is much more efficient and better suited for these experiments. Future work will investigate the ability of spheroid-cultured ADSCs to differentiate into particular cell lineages, with an emphasis on skeletal myogenic differentiation.

Previously Presented/Performed?

Fourth Annual Showcase of Undergraduate Research and Creative Endeavors (SOURCE), Winthrop University, April 2018

Grant Support?

Supported by SC INBRE and INBRE Developmental Research Project grants from the National Institute of General Medical Sciences (NIH-NIGMS)

Start Date

20-4-2018 2:15 PM

End Date

20-4-2018 4:15 PM

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Apr 20th, 2:15 PM Apr 20th, 4:15 PM

Spheroid Culture of Murine and Human Adipose-Derived Stem Cells Alters the Expression of Genes that Regulate Developmental Potency

Richardson Ballroom (DIGS)

Adipose-derived stem cells (ADSCs) are multipotent, mesenchymal stem cells that are found within the microvasculature of adipose tissue. While ADSCs have the potential to differentiate into multiple cell lineages, they cannot match the differentiation potential of pluripotent stem cells, such as embryonic stem cells or induced pluripotent stem cells. Previous research has shown that stem cells are sensitive to the composition and dimensionality of their culture environment. We have shown that culturing murine ADSCs as three-dimensional spheroids alters their gene expression compared to ADSCs cultured using traditional, two-dimensional methodology. We hypothesized that human ADSCs cultured as spheroids and in two dimensions would also exhibit differential gene expression, and that the changes in gene expression would mirror those observed in murine ADSCs. To investigate our hypothesis and optimize our protocol for generating spheroids, we compared our traditional hanging-drop method to the use of micropore plates that are specially designed for producing spheroids. Our results suggest that human ADSC gene expression is significantly altered by spheroid culture, with some changes mirroring what was observed in murine ADSCs. The method of spheroid generation had little impact on the results; however, the workflow associated with use of the micropore plates is much more efficient and better suited for these experiments. Future work will investigate the ability of spheroid-cultured ADSCs to differentiate into particular cell lineages, with an emphasis on skeletal myogenic differentiation.