MacFarlane or Macfarlane is a surname shared by:
- Alan Macfarlane (born 1941), a professor of anthropological science at Cambridge University
- Alexander Macfarlane (1851-1913), a Scottish-Canadian logician, physicist, and mathematician
- Alexander Macfarlane (politician) (1818-1898), a Nova Scotia lawyer and politician
- Ian Macfarlane (economist), economist and former Governor of the Reserve Bank of Australia
- Jean Macfarlane, American psychologist
- John MacFarlane, the founder and CEO of Sonos, which makes digital audio equipment
- John Edward Macfarlane, the editor of Toronto Life magazine
- John Muirhead Macfarlane (1855–1943), a Scottish botanist and professor
- Laura Macfarlane, rock and roll musician
- Luke Macfarlane (b. 1980), Canadian actor
- Malcolm MacFarlane (1853–1931), a Scottish Gaelic scholar and songwriter.
- Neil Macfarlane, (b. 1936) a British Conservative politician and former Minister for Sport
- Norman Macfarlane, Baron Macfarlane of Bearsden, (b. 1926) a British Conservative member of the House of Lords
- Rachael MacFarlane (born 1976), a voice actress and sister of Seth MacFarlane
- Robert Macfarlane (born 1976), a British travel writer, cultural historian, and literary critic
- Robert MacFarlane (1835-1872), a lawyer and Liberal member of the Canadian House of Commons
- Robert Macfarlane (New Zealand) (1900 - 1982), a New Zealand politician
- Robert Gwyn Macfarlane, hæmatologist
- Seth MacFarlane (born 1973), an American animator and voice actor, best known for his show Family Guy
- Todd MacFarlane (born 1961), a Canadian cartoonist, best known for his comic Spawn
- Macfarlane, Lang & Co., Ltd, a biscuit firm which merged with McVitie & Price to form United Biscuits