Spheroid Culture of Human Adipose Derived Stem Cells to Alter Regenerative Potential
Poster Number
006
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 reside in the microvasculature of adipose tissue. While they are partially defined by their ability to differentiate into multiple cell lineages, their directed differentiation into many lineages of interest remains inefficient. Our lab is specifically interested in the ability of ADSCs to differentiate into skeletal myocytes. The goal of this project is to differentiate ADSCs into skeletal myocytes more efficiently, by first moving the cells to a more developmentally potent state. Multiple factors have shown promise in enhancing cells’ regenerative potential. Specifically, three-dimensional spheroid culture is known for its ability to enhance cells’ differentiation potential. We hypothesized that ADSC spheroids generated using micropore well inserts would show changes in global gene expression indicative of a more developmentally potent state, and that the addition of 5-azacytidine, a DNA methylation-blocking analogue of cytidine, would provide a synergistic enhancement of the effects of spheroid culture. To test our hypothesis, we generated ADSC spheroids and compared the transcriptomes of spheroid-cultured ADSCs to ADSCs cultured using the traditional two-dimensional method with or without the addition of 5-azacytidine. RNA sequencing results showed differences in the gene expression profiles of all of the experimental and control groups. The expression profile of the plated spheroids was more similar to two-dimensionally cultured cells than to three-dimensional spheroids. The large amount of data generated requires further analysis to determine the relevance of differentially expressed genes to developmental potency and directed differentiation into skeletal myocytes.
Grant Support?
Supported by SC INBRE and INBRE Developmental Research Project grants from the National Institute of General Medical Sciences (NIH-NIGMS)
Start Date
12-4-2019 12:00 PM
End Date
April 2019
Spheroid Culture of Human Adipose Derived Stem Cells to Alter Regenerative Potential
Rutledge Building
Adipose Derived Stem Cells (ADSCs) are multipotent mesenchymal stem cells that reside in the microvasculature of adipose tissue. While they are partially defined by their ability to differentiate into multiple cell lineages, their directed differentiation into many lineages of interest remains inefficient. Our lab is specifically interested in the ability of ADSCs to differentiate into skeletal myocytes. The goal of this project is to differentiate ADSCs into skeletal myocytes more efficiently, by first moving the cells to a more developmentally potent state. Multiple factors have shown promise in enhancing cells’ regenerative potential. Specifically, three-dimensional spheroid culture is known for its ability to enhance cells’ differentiation potential. We hypothesized that ADSC spheroids generated using micropore well inserts would show changes in global gene expression indicative of a more developmentally potent state, and that the addition of 5-azacytidine, a DNA methylation-blocking analogue of cytidine, would provide a synergistic enhancement of the effects of spheroid culture. To test our hypothesis, we generated ADSC spheroids and compared the transcriptomes of spheroid-cultured ADSCs to ADSCs cultured using the traditional two-dimensional method with or without the addition of 5-azacytidine. RNA sequencing results showed differences in the gene expression profiles of all of the experimental and control groups. The expression profile of the plated spheroids was more similar to two-dimensionally cultured cells than to three-dimensional spheroids. The large amount of data generated requires further analysis to determine the relevance of differentially expressed genes to developmental potency and directed differentiation into skeletal myocytes.
