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A native of North Charleston, SC, Charlene Gates Stuart received her Bachelor of Science Degree in Accounting from the University of South Carolina in 1968. She joined the Medical College of South Carolina in 1968 as Assistant to the Budget Director. In 1977 Stuart obtained her Masters in Business Administration from The Citadel.

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From 1968 until 1981, Stuart held increasingly responsible financial management positions. In April of 1982, while serving as MUSC Controller and Assistant Treasurer, Stuart was appointed the Associate Administrator for External Affairs in the Health Care Financing Administration/US Department of Health and Human Services by President Ronald Reagan. She was named the first female Executive Director of the MUSC Medical Center in July of 1987. MUSC President James B. Edwards, DMD, appointed Stuart MUSC Vice President of Finance & Administration in July 1991. In 1994 after re-engineering the Medical Center, Stuart was named to the newly created position of Vice President for Clinical Operations, MUSC and CEO, MUSC Medical Center. In 1997 the MUSC Medical Center was recognized for quality of care, efficiency of operations, and sustainability over time in the "100 Top Hospitals Benchmarks for Success". Also in 1997, Stuart was honored by the South Carolina General Assembly with an appreciation resolution for her outstanding service to MUSC and the State of South Carolina. That year Stuart moved to Saudi Arabia with her husband where she was named the first female Senior Administrator at the King Faisal Specialist Hospital and Research Centre in Riyadh.

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While living in Riyadh, Stuart was diagnosed with acute myeloid leukemia. In 2000 she received a successful stem cell transplant at the Hollings Cancer Center. Currently she is enjoying life at home in Charleston with her spouse Robert K. Stuart, MD, and her grandchildren Darren, Gates, and Alexandra.