The present study contributes elements to the literature for the understanding of those institutional factors of organizations that promote the creation of spin-offs in universities. Institutional barriers to spin-off creation within universities are addressed from a qualitative perspective. The study was carried out in Colombia, a country with low levels of innovation and entrepreneurship. Purposive and snowball sampling were employed to identify experts. Participants were selected in accordance with their extensive experience in entrepreneurship, innovation, or technology transfer. The sample size was decided by saturation criteria, and 15 interviews were selected with this method. The semi-structured interview was chosen for information collection, and a thematic analysis was employed for data interpretation. Several aspects which permit suggestions for the future spin-off research agenda were identified: perceptions of the university, teaching focus, academic cultures that reward publication as a means of knowledge transfer, restrictions for public university spin-offs in developing countries, few economic and non-economic incentives for entrepreneurship, trust as a determining cultural characteristic, and limitations to the understanding of the spin-off concept that affect their creation. These results may be extrapolated to Latin America and other developing regions with similar entrepreneurial ecosystems.