Van Sinderen Lindsley was born in Greensboro, North Carolina, in 1840. His brother, Philip Lindsley, D.D., was founder and president of the University of Nashville. Primarily educated in Greensboro, Lindsley graduated from the University of Nashville in 1861 and received his M.D. from there later as well. Upon receiving his medical degree, he began an appointment in the Anatomy Department at his alma mater, which he held until 1868.

Upon returning from a tour of professional observation through some of the principal hospitals in Europe and the major cities of the United States, Lindsley was appointed Chair of Surgical Anatomy at the University of Nashville, which he occupied until 1871, when he was elected to the Chair of Physiology. This department also taught diseases of the eye and ear.

Relinquishing general practice, in which he had been engaged at Nashville for a dozen years, Lindsley adopted otolaryngology as his specialty. He also held official appointments as President of the Nashville Medical Society and as member of the American Association for the Advancement of Science. Lindsley was a delegate from the State Medical Society to the International Medical Congress held in Philadelphia in 1876.