The Honorable Donetta W. Ambrose  was born on November 5, 1945 in New Kensington, Pennsylvania, the daughter of Chester J. and Mary Groza Wypiski. She married J. Raymond Ambrose, Jr. on August 19, 1972. They have one son, J. Raymond Ambrose, III. She graduated from Arnold High School in 1963, Duquesne University in 1967, with the degree of Bachelor of Arts, and was awarded a J.D. cum laude at the Duquesne University School of Law in 1970. She was admitted to practice before the Supreme Court of Pennsylvania in December 1970 and is a member of the Westmoreland County, Allegheny County, Pennsylvania, American and Women's Bar Associations. She is also a Fellow of the American Bar Foundation, a member of the American Judicature Society, the National Association of Women Judges, and the Inns of Court.

She was a law clerk to the late Pennsylvania Supreme Court Justice Louis L. Manderino from 1970 to 1972, an Assistant Attorney General for the Commonwealth of Pennsylvania from 1972 to 1974, an Assistant District Attorney for Westmoreland County from 1977 to 1982 and engaged in the general practice of law from 1974 to 1982. She was elected to the Westmoreland County Court of Common Pleas in November of 1981 on which she served from January 1982 until her appointment to the United States District Court for the Western District of Pennsylvania on November 24, 1993.

She has been active in the Pennsylvania Commission for Women in the Profession, Big Brothers and Sisters of Westmoreland County and various civic, religious and charitable activities. She resides with her family in Lower Burrell, Pennsylvania.

Judge Ambrose was the first female Chief Judge for the Western District of Pennsylvania.  She served as Chief Judge from 2002 - 2009.