Grounded on the research-established advantages of Lesson Study (LS) on the professional development of teachers in other countries as well as in the basic education in the Philippines, a study aimed at establishing the impact of Lesson Study on the practices, pedagogical content knowledge (PCK) and technological pedagogical content knowledge (TPCK) of two College Biology teachers was conducted. Following the design-based research, the effect of the artifact developed by the LS- participating teachers constitutes LS- generated teaching materials which included one Lesson Guide for each lesson on the attitude and cognitive skills of 57 college Biology students was also determined. Findings show that LS improved the practices and self-evaluated PCK and TPCK, and student-rated subject matter knowledge and TPCK of teachers whereas LS-generated teaching materials significantly increase the cognitive skills and improve the attitudes of students toward Biology. In conclusion, participating in three cycles of LS contributed to better preparation of teachers for their classes, their PCK and TPCK and these were translated to enhanced cognitive skills and attitude of students. A professional development model that is able to address the challenges of adopting LS in the College is also proposed.