In March 1825, the first class of forty students, guided by nine professors, matriculated at the University of Virginia. At this time, the duties of enslaved laborers began to shift from construction to the maintenance of the ten pavilions, fifty-four student rooms, and Rotunda (then still under construction), as well as the care of the people who used them. In 1824, the board of visitors had prohibited incoming students, but not faculty, from bringing slaves they owned onto Grounds. This rule contrasted with the policies of other Virginia colleges, including the College of William and Mary in Williamsburg , where students often brought their personal slaves to wait on them. In 1840, the university even prohibited students who lived off-Grounds from keeping slaves. In 1826, as part of an attempt to keep unwanted slaves and free blacks from Grounds, the faculty instructed the university proctor to license all slaves owned and hired by the university. Enslaved men and women were required to wear the licenses on their persons. In 1829, the university instituted a regular slave patrol and continued to employ an overseer until at least 1846.

In the years before the Civil War, more than 100 enslaved men and women worked at the university at any given time. Many of them labored in the hotels, or boardinghouses, located on the Academical Village's East and West Ranges. For an annual fee of between $150 and $165, hotelkeepers provided students with three daily meals, as well as furniture, linens, firewood, ice, water, and laundry and cleaning services. Slaves owned by the hotelkeepers—and not by the university—performed all associated tasks.

In 1842, the faculty articulated thirteen specific chores to be performed by the hotelkeepers' slaves: fetching water and clean towels, making fires in winter, cleaning rooms, making beds, cleaning candlesticks, washing fireplaces (once a week), blacking andirons (once a week), carrying water to each dormitory (twice daily), washing windows (once a month), whitewashing fireplaces (twice a summer), blacking students' shoes, carrying ice, and stacking wood. Between two forty-five and three o'clock every afternoon, one slave from each hotel was made available to run errands for students. In addition, hotel slaves cooked the day's meals and cleaned up afterward. On average, there was one slave for every twenty students.

Other slaves were owned or, more rarely, hired by the university. Anatomical Lewis, possibly owned by the university as early as 1830 and as late as 1860, assisted the anatomy professor in what the historian Catherine S. Neale has described as "a sordid job and poor living conditions," making him "an outcast of the community." The enslaved man Lewis Commodore had been hired out to the university for a number of years when school officials, learning that his master intended to sell him, purchased him on July 18, 1832, for $580. Commodore rang the Rotunda's so-called Medway Bell, which marked the day's schedule, until 1847, when the duty passed to Henry Martin, a free black. Beginning in 1834, Commodore also opened the library each morning and tended to lecture rooms, ensuring they were tidy and warm. In 1851, the faculty determined that he had "repeatedly and grossly neglected his duties," a dereliction possibly related to his reported abuse of alcohol, and relieved Commodore of his responsibilities. He was then sold.

Enslaved workers lived in various makeshift quarters. Until 1834, when slaves were prohibited from residing in the Rotunda, Lewis Commodore likely lived in a room on that building's ground floor. A brick building near the gates of the university housed slaves until 1838, when it was appropriated for an infirmary. Faculty-owned slaves often lived in the basements of the pavilions or in unclaimed rooms, while others, including Anatomical Lewis, may have occupied structures built in the university's gardens. Hotel slaves likely found space in their respective hotels.

The quality and timeliness of the hotel slaves' service were often the subject of complaints by students and faculty. Slaves were charged with neglecting to clean rooms and not completing their work quickly or subserviently enough. In some cases, hotelkeepers responded by blaming their slaves' poor health, which may have been due to poor diet and living conditions. Many slaves also may have had more work than they could accomplish in a timely manner. The historian Philip Alexander Bruce, writing in 1920 but likely reflecting attitudes from the previous century, blamed in part "slipshod slave service" for frequent outbreaks of disease, including typhoid fever, especially on the East Range: "The inefficient services of that day furnished by lazy, untrained slaves, who had been hired out only too often because their characters were bad, was an additional cause of these outbursts of distemper." School administrators generally settled such complaints by instructing hotelkeepers to supervise their enslaved laborers more closely.