Telemedicine is both effective and able to provide efficient care at a lower cost. It also enjoys a high degree of acceptance among users. The Technology Acceptance Model proposed is based on the two main concepts of ease of use and perceived usefulness and is comprised of three dimensions: the individual context, the technological context and the implementation or organizational context. At present, no short, validated questionnaire exists in Catalonia to evaluate the acceptance of telemedicine services amongst healthcare professionals using a technology acceptance model. This article aims to statistically validate the Catalan version of the EU project Health Optimum telemedicine acceptance questionnaire. The study included the following phases: adaptation and translation of the questionnaire into Catalan and psychometric validation with construct (exploratory factor analysis), consistency (Cronbach’s alpha) and stability (test–retest) analysis. After deleting incomplete responses, calculations were made using 33 participants. The internal consistency measured with the Cronbach’s alpha coefficient was good with an alpha coefficient of 0.84 (95%, CI: 0.79–0.84). The intraclass correlation coefficient was 0.93 (95% CI: 0.852–0.964). The Kaiser–Meyer–Olkin test of sampling showed to be adequate (KMO = 0.818) and the Bartlett test of sphericity was significant (Chi-square 424.188; gl = 28; p < 0.001). The questionnaire had two dimensions which accounted for 61.2% of the total variance: quality and technical difficulties relating to telemedicine. The findings of this study suggest that the validated questionnaire has robust statistical features that make it a good predictive model of healthcare professional’s satisfaction with telemedicine programs.