Peale was born in 1741 in Chester, Queen Anne's County, Maryland, the son of Charles Peale and his wife Margaret. He had a younger brother, James Peale (1749-1831). Charles became an apprentice to a saddle maker when he was thirteen years old. Upon reaching maturity, he opened his own saddle shop; however, when his Loyalist creditors discovered he had joined the Sons of Liberty, they conspired to bankrupt his business.

In 1762, Peale married Rachel Brewer (1744-1790), who bore him ten children, most named for Peale's favorite artists, male and female. The sons included Raphaelle Peale (1774-1825), Rembrandt Peale (1778-1860), who was another famous portrait painter and museum owner/operator in Baltimore, and scientific inventor and businessman, Titian Peale I (1780-1798), and Rubens Peale (1784-1865). Among the daughters: Angelica Kauffman Peale (named for Angelica Kauffman, Peale's favorite female painter) married Alexander Robinson, her daughter Priscilla Peale wed Dr. Henry Boteler, and Sophonisba Angusciola Peale (named for Sofonisba Anguissola) married Coleman Sellers.  After Rachel's death in 1790, Peale married Elizabeth de Peyster (d. 1804) the next year. With his second wife, he had six additional children. One son, Franklin Peale, born on October 15, 1795, became the Chief Coiner at the Philadelphia Mint. Their youngest son, Titian Ramsay Peale (1799-1885), became an important naturalist and pioneer in photography. Their daughter, Elizabeth De Peyster Peale (1802-57), married William Augustus Patterson (1792-1833) in 1820.  Hannah More, a Quaker from Philadelphia, married Peale in 1804, becoming his third wife. She helped raise the younger children from his previous two marriages.  Peale's slave, Moses Williams, was also trained in the arts while growing up in the Peale household and later became a professional silhouette artist.  In 1810, Peale purchased a farm in Germantown where he intended to retire. He named this estate 'Belfield', and cultivated extensive gardens there. After Hannah's death in 1821, Peale lived with his son Rubens and sold Belfield in 1826. Peale died on February 22, 1827, and was buried at the Saint Peter's Episcopal Church in Philadelphia.

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