The first administrator and teacher at Mount Holyoke was Mary Lyon, the founder of the school who was Principal from 1837 until her death in 1849 as well as a teacher in chemistry classes. Initially, she hired Eunice Caldwell as Associate Principal with Amanda A. Hodgman and Mary W. Smith as additional teachers. Lyon also relied on founding members of Mount Holyoke's Board of Trustees to assist her with managing the institution's finances and supervising the construction, furnishing, and maintenance of the Seminary Building where students and teachers lived and worked. She soon began hiring Mount Holyoke graduates as teachers at the school, a practice that continued for much of the nineteenth century. Teachers usually taught a variety of subjects and often supervised the domestic work system Lyon established whereby students did most of the cooking, cleaning, and laundry at the school (thus keeping the tuition rate low by eliminating the need for a paid staff). Men were hired to perform heavy manual labor and a Steward was responsible for the overall maintenance and upkeep of buildings and grounds. Trustees continued to act as Treasurers of Mount Holyoke until the early twentieth century. Other early staff members included a librarian (Mary O. Nutting, Class of 1852) and the Director of the Observatory (Elisabeth Bardwell, Class of 1866).

Teachers began specializing in particular subjects in the late 1880s. Elizabeth Mead, President of Mount Holyoke from 1890-1900, required teachers to earn a Ph.D. in their respective fields and by 1896 faculty were organized into a formal group which held regular meetings and actively shaped the College's curriculum. The first male teachers, Asa Kinney (botany) and William Churchill Hammond (music), were hired in 1899. When Mount Holyoke began a graduate program in the 1890s, graduate students (including those from countries other than the United States) began working as teaching or laboratory assistants at the College. The first paid sabbaticals were granted to faculty in 1925. Mount Holyoke's faculty has steadily become increasingly diverse in response to changes in the curriculum and the College's affirmative action initiatives.

The College has created many staff positions to meet the growth and changing needs of the school. For example, the first Registrar, Caroline Boardman Greene (Class of 1889) was appointed in 1893 and Florence Purington (Class of 1886) became the first Dean of the College in 1907. When students began living in separate dormitories after the destruction of the Seminary Building by fire in 1896, house mothers were hired to oversee students' activities in each residence. Maids and other staff were hired in increasing numbers after the domestic work system ended in 1914. Other staff members have been appointed as needed, such as those who support the technological needs of faculty and staff as employees of Library, Information, and Technology Services.