The Use of Magnetic Cell Sorting to Obtain Multilineage-Differentiating Stress Enduring Cells from Human Adipose Derived Stem Cells
College
College of Arts and Sciences
Department
Biology
Abstract
Multilineage-differentiating stress enduring (Muse) cells are a unique subpopulation of mesenchymal stem cells (MSC) that are pluripotent and have the ability to self-renew for multiple generations. Muse cells also have an advanced ability to receive damage signals, survive in stress-filled environments, and exhibit low tumorigenic activity. Due to their expression of the cell surface antigen SSEA-3, Muse cells in a heterogeneous population of MSCs can be separated from non-Muse cells. We hypothesized that human adipose derived stem mesenchymal cells (ADSCs) can be sorted into Muse and non-Muse populations on the basis of SSEA-3 expression using a magnetic cell sorting strategy. To test our hypothesis, cells were magenetically sorted, and the expression of genes associated with enhanced developmental potency was compared between Muse and non-Muse populations of ADSCs using real-time PCR. Our working hypothesis is that Muse ADSCs will exhibit significantly greater levels of expression of genes associated with enhanced developmental potency than non-Muse ADSCs. In addition to gene expression studies, both Muse and non-Muse cells were plated on poly-HEMA plates to assess their ability to form M-clusters – a known characteristic of Muse cells. Future work will include sorting human ADSCs into Muse and non-Muse populations using fluorescence activated cell sorting (FACS), as well as testing the developmental potential of Muse cells in three-dimensional culture systems.
Honors Thesis Committee
Matthew Stern, Ph.D.; Laura Glasscock, Ph.D.; and Courtney Guenther, Ph.D.
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 3:30 PM
The Use of Magnetic Cell Sorting to Obtain Multilineage-Differentiating Stress Enduring Cells from Human Adipose Derived Stem Cells
West 221
Multilineage-differentiating stress enduring (Muse) cells are a unique subpopulation of mesenchymal stem cells (MSC) that are pluripotent and have the ability to self-renew for multiple generations. Muse cells also have an advanced ability to receive damage signals, survive in stress-filled environments, and exhibit low tumorigenic activity. Due to their expression of the cell surface antigen SSEA-3, Muse cells in a heterogeneous population of MSCs can be separated from non-Muse cells. We hypothesized that human adipose derived stem mesenchymal cells (ADSCs) can be sorted into Muse and non-Muse populations on the basis of SSEA-3 expression using a magnetic cell sorting strategy. To test our hypothesis, cells were magenetically sorted, and the expression of genes associated with enhanced developmental potency was compared between Muse and non-Muse populations of ADSCs using real-time PCR. Our working hypothesis is that Muse ADSCs will exhibit significantly greater levels of expression of genes associated with enhanced developmental potency than non-Muse ADSCs. In addition to gene expression studies, both Muse and non-Muse cells were plated on poly-HEMA plates to assess their ability to form M-clusters – a known characteristic of Muse cells. Future work will include sorting human ADSCs into Muse and non-Muse populations using fluorescence activated cell sorting (FACS), as well as testing the developmental potential of Muse cells in three-dimensional culture systems.
