Born in Geneva, Switzerland. Spent ten years as a portrait painter in the West Indies. Arrived in New York City in 1765 and settled in Philadelphia in 1766. He was drafted into the Pennsylvania militia in 1777 but petitioned for immunity as a foreigner, even though he had become a naturalized citizen in 1769.

He served as curator for the American Philosophical Society 1776 to 1781. In 1782 he advertised for his "American Museum" in Philadelphia. It is considered the second museum in the United States. He amassed a collection of coins of the world numbering hundreds of pieces for his museum. Responsible for the earliest known sale of a coin collection by private treaty in America (1772), and by auction (1785). Never married. He died in Philadelphia.

On February 1, 1779, he drew a profile of George Washington from life. The drawing was engraved and sold to the public. It became the source for the portrait of Washington on many medals. Attributing these exactly has been the subject of some conjecture and disagreement among Washington scholars. Breen and Swiatek refer to both Du Simitiere and Houdon as sources for the portrait of Washington on the Lafayette Silver Dollar.