Context: As industries are heading for digital transformation through Industry 4.0, the concept of Digital Twin (DT) - a software for digital transformation, has become popular. Many industries use DT for its advantages, such as predictive maintenance and real-time remote monitoring. Within DT domain, an emerging topic is the concept of an ecosystem—a digital platform that would create value for different stakeholders in an ecosystem of DT-driven products and services. The identification of potential stakeholders and their requirements provides valuable contributions to the development of healthy Digital Twin Ecosystems (DTE). However, current empirical knowledge of potential stakeholders and their requirements are limited. Objective/Methodology: Thus, the objective of this research was to explore potential stakeholders and their requirements. The research employed an empirical research methodology in which semi-structured interviews were conducted with DT professionals for data collection. Results: Data analysis of the study revealed 13 potential stakeholders who were categorized as primary (manufacturers, suppliers, subcontractors, and intelligent robots), secondary (maintenance service providers, platform integration service providers, tech companies, etc.), and tertiary (research organizations, third-party value-added service providers, cyber security firms, etc.). This study also presents the different requirements of these stakeholders in detail. Contribution: The study contributes to both research and industry by identifying possible stakeholders and their requirements. It contributes to the literature by adding new knowledge on DTEs and fills a research gap while contributing industry by providing ample knowledge to the industry’s practitioners that is useful in the development and maintenance of a healthy DTE.