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This study explores the development of a teacher’s technological pedagogical content knowledge (TPCK) (Koehler et al, 2013) during his differentiated instruction (DI) (Tomlinson, 2001) practices in the English language classrooms of a three-year school-university collaboration project. A longitudinal qualitative case study was conducted with data collected through questionnaires, interviews, observations and documents and artefacts to investigate the changes in the teacher’s TPCK and some potential personal and contextual factors affecting the dynamic interactions between TPCK core components and the DI elements. Research findings enrich the TPCK framework and shed light on DI teacher professional development. Implications for application of technology-facilitated learning in DI practices are also discussed.