Information Technology (IT) application has become popular in the modern generation. Almost every enterprise establishes the information system to improve its production or service depending on its organization needs. It has been a worldwide focus on applying IT to the learning domain, and a principal educational policy on integrating IT with teaching. Many schools have been integrated into an e-Portfolio which has been used as the tool of teaching innovation in teaching and evaluation currently. DeLone & McLean (1992) proposed "Information System Successful Model, ISSM", which provided the research direction of the academic circles information system and the consideration factor of the success key. Davis's "Technology Acceptance Model" (TAM) receives scholarly studies and the practical realm for public figure's widespread application in various kinds of technology areas, and pass through many real diagnoses during these years, and the results all pointed out that TAM is a stable tool. The main purpose of this research is to carry on investigating analysis and to inquire into student's usage attitude, user's satisfaction and usage will for "e-Portfolio" based on ISSM and TAM. According to the research object, students of different back ground use e-Portfolio, which holds discrepancy and correlation of student's usage attitude. The questionnaire was designed based on research model and the proposed hypotheses. According to this study and the results of the questionnaire, some conclusions are suggested as follows: 1. Using attitude of the students toward the "e-Portfolio" influences their willingness to use it. The result corresponded with theory of "TAM". 2. Users' satisfaction of the students about the "e-Portfolio" influences their willingness to use it. The result corresponded with theory of "ISSM". 3. The students of different characters are different from using attitudes, satisfaction and users' acceptance. The results showed that the University students with external factors conform to the TAM and ISSM. These results also improved our understanding of the external factors affecting the acceptance of e-Portfolio, which could be help development and popular usage of e-Portfolio in the school and make teacher and student reach the reference of the object of e-Portfolio application of the learning process.