The Middlesex and London Region has a rich cultural heritage and is home for immigrants from around the globe. The predominant language is English but almost 19% of the population speak something other than English or French as their first language.

| Mother tongue | Middlesex & London Region | ||
| Total | Male | Female | |
| Total population | 417,670 | 202,665 | 215,005 |
| English only | 332,685 | 162,020 | 170,665 |
| French only | 5,660 | 2,485 | 3,170 |
| English and French | 675 | 325 | 350 |
| Other language(s) | 78,655 | 37,830 | 40,820 |
| Source: Statistics Canada 2006 Census | |||
|
Knowledge of official languages |
Middlesex & London Region | ||
| Total | Male | Female | |
| Total population | 417,670 | 202,665 | 215,005 |
| English only | 382,895 | 188,140 | 194,755 |
| French only | 275 | 145 | 130 |
| English and French | 29,760 | 12,625 | 17,140 |
| Neither English nor French | 4,735 | 1,755 | 2,980 |
| Source: Statistics Canada 2006 Census | |||
|
Language spoken |
Middlesex & London Region | ||
| Total | Male | Female | |
| Total population | 417,670 | 202,665 | 215,010 |
| English | 373,120 | 181,470 | 191,655 |
| French | 1,745 | 785 | 960 |
| Non-official language | 37,605 | 17,865 | 19,735 |
| English and French | 450 | 225 | 225 |
| English and non-official language | 4,645 | 2,260 | 2,385 |
| French and non-official language | 40 | 25 | 15 |
| English, French and non-official language | 55 | 30 | 25 |
| Source: Statistics Canada 2006 Census | |||
| Immigrant status and period of immigration | Middlesex & London Region | ||
| Total | Male | Female | |
| Total population | 417,670 | 202,665 | 215,005 |
| Non-immigrants | 330,030 | 160,580 | 169,445 |
| Immigrants | 83,450 | 39,840 | 43,610 |
| Before 1991 | 53,645 | 25,610 | 28,030 |
| 1991 to 2000 | 17,280 | 8,195 | 9,080 |
| 2001 to 2006 | 12,530 | 6,035 | 6,500 |
| Non-permanent residents | 4,190 | 2,245 | 1,945 |
| Source: Statistics Canada 2006 Census | |||
| Citizenship | Middlesex & London Region | ||
| Total | Male | Female | |
| Total population | 417,670 | 202,665 | 215,005 |
| Canadian citizens | 395,715 | 191,930 | 203,785 |
| Canadian citizens under age 18 | 88,465 | 45,950 | 42,515 |
| Canadian citizens age 18 and over | 307,245 | 145,980 | 161,270 |
| Not Canadian citizens | 21,955 | 10,735 | 11,225 |
| Source: Statistics Canada 2006 Census | |||