Matoušek (feminine Matoušková) is a Czech surname. Notable people with the surname include:
- Bohuslav Matoušek (born 1949) Czech violinist
- Gabriela Matoušková, Czech footballer
- Iveta Matoušková, Czech handballer
- Jan Matoušek, Czech footballer
- Jan Matoušek (rower), Czech rower
- Jaroslav Matoušek, Czech athlete
- Jiří Matoušek (disambiguation), multiple people
- Josef Matoušek (historian), Czech historian
- Josef Matoušek (athlete), Czech athlete
- Karel Matoušek, Czech wrestler
- Kateřina Matoušková, Czech-Canadian figure skater
- Matylda Matoušková-Šínová, Czech gymnast
- Petr Matoušek, Czech cyclist
- Tomáš Matoušek, Slovak ice hockey player