According to Official Guinness Records,
The most players in a 24-hour Guitar Hero session is 594, achieved by Activision Italy and gamers in the city of Verona, Italy as part of the launch of Guitar Hero 5 from 11-12 September 2009.
Sweden came a close second, with Madrid and Utrecht also doing well in the multi venue organised attempt on this record.
For a complete list of 2009 records, please visit 2009 Guinness Records in Italy.