Inducing Myogenic Differentiation of Human Adipose-Derived Stem Cells through Culture with Epigenetic Modifier 5-Azacytidine
College
College of Arts and Sciences
Department
Biology
Abstract
Stem cells are undifferentiated cells that have the capability to differentiate into one or more cell lineages. Adipose-derived stem cells (ADSCs) are multipotent, mesenchymal stem cells that are located within the microvasculature of adipose tissue. Although ADSCs have the ability to differentiate into multiple cell lineages, there has been little documented success in differentiating ADSCs into myogenic lineages, specifically skeletal muscle. Many of the reports of skeletal myogenic differentiation of ADSCs have relied on a protocol that includes treatment with the DNA methylation inhibitor 5-azacytidine. In addition, previous work in our lab suggests that culture with 5-azacytidine induces the expression of genes associated with enhanced developmental potency. Thus, we hypothesized that a myogenic induction protocol that includes treatment with 5-azacytidine in combination with horse serum and low-glucose DMEM would result in the myogenic differentiation of ADSCs into skeletal myoblasts and myotubes. To test our hypothesis, we employed real-time PCR to monitor changes in the expression of genes that regulate myogenic differentiation and immunofluorescent staining to detect the appearance of myotubes. Future work will include replicating this experiment with ADSCs grown in/on a variety of substrates that are expected to further enhance myogenic differentiation. The ability to induce myogenic differentiation of ADSCs would allow ADSCs to serve as a plentiful source of myogenic cells for skeletal muscle tissue engineering and regenerative medicine applications.
Honors Thesis Committee
Matthew Stern, Ph.D.; Courtney Guenther, Ph.D.; and Kristi Westover, 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 1:00 PM
Inducing Myogenic Differentiation of Human Adipose-Derived Stem Cells through Culture with Epigenetic Modifier 5-Azacytidine
West 221
Stem cells are undifferentiated cells that have the capability to differentiate into one or more cell lineages. Adipose-derived stem cells (ADSCs) are multipotent, mesenchymal stem cells that are located within the microvasculature of adipose tissue. Although ADSCs have the ability to differentiate into multiple cell lineages, there has been little documented success in differentiating ADSCs into myogenic lineages, specifically skeletal muscle. Many of the reports of skeletal myogenic differentiation of ADSCs have relied on a protocol that includes treatment with the DNA methylation inhibitor 5-azacytidine. In addition, previous work in our lab suggests that culture with 5-azacytidine induces the expression of genes associated with enhanced developmental potency. Thus, we hypothesized that a myogenic induction protocol that includes treatment with 5-azacytidine in combination with horse serum and low-glucose DMEM would result in the myogenic differentiation of ADSCs into skeletal myoblasts and myotubes. To test our hypothesis, we employed real-time PCR to monitor changes in the expression of genes that regulate myogenic differentiation and immunofluorescent staining to detect the appearance of myotubes. Future work will include replicating this experiment with ADSCs grown in/on a variety of substrates that are expected to further enhance myogenic differentiation. The ability to induce myogenic differentiation of ADSCs would allow ADSCs to serve as a plentiful source of myogenic cells for skeletal muscle tissue engineering and regenerative medicine applications.
