Thousands of people joined the event, held each year to mark the anniversary of the Supreme Court decision legalising abortion.
Vice-President Mike Pence told them: "Life is winning again in America".
It comes less than a week after a large protest against Mr Trump was held in the nation's capital city.
Jeanne Mancini, president of the March for Life, says that 58 million abortions have been performed in the US since the Supreme Court legalised the practice in 1973.
"We don't want to judge them. We want to help them," said Joi Hulecki, 63, a nurse practitioner from Orlando, Florida.
"We consider ourselves pro-women too," she added.
An estimated 500,000 people protested for women's rights last Saturday - including many supporters of abortion rights - at the Women's March.
Mr Pence is the first president or vice-president ever to attend the rally
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