This paper investigates the triple helix (industry, knowledge workers and governments) cooperation on knowledge co-production and valorisation for innovation, which took place in seven horticultural regions in the Netherlands. It thus provides more empirical insight into the functioning of this form of cooperation. Based on a secondary multiple case study analysis, this paper sets out to ascertain what enabled triple helix cooperation in the seven regions with respect to the organisation, the formulation and support for goals and action on knowledge co-production and valorisation. The results indicate that in order to stimulate innovation through triple helix cooperation, the different partners first need to build a proper working relationship and a common language. In order to accomplish this, primary aims for innovation should not be formulated too ambitiously (i.e. too far beyond the entrepreneurs’ daily practice, in particular SMEs). Knowledge workers and policy makers often want to stimulate knowledge co-production and valorisation more radically and quickly. Hence, they have to temper their ambitions. Procedures regarding the cooperation should be rather simple and flexible. Once a steady working relationship and a common language are developed, then the triple helix collaboration can focus on taking the innovation ambition to a higher level in order to realise more valuable change. At first, entrepreneurs have to experience how they can profit from the cooperation and learn to incorporate knowledge co-production and valorisation step-by-step in their business strategy, including financial investments.