Achieving truly dynamic virtual organisations (VOs) as short-term, goal-oriented and opportunity-based collaborative networked organisations (CNOs), in response to emerging collaboration (business) opportunities, requires the pre-existence of proper 'breeding environments' enabling and providing the necessary conditions and mechanisms for the rapid and fluid configuration of VOs. VO breeding environments (VBEs) act as long-term supporting networks underlying the nesting environments for preparing their members and maintaining this preparedness over the time for their effective and ad-hoc collaboration in VOs. This article introduces a VBE reference model as a reference guide for VBE initiators, VBE managers and CNO experts responsible for the creation and operation of breeding environments in different domains and application environments. As part of the VBE reference model, a VBE management framework is also introduced as a set of management activities and supporting tools to ensure that a VBE can fulfil all tasks required to achieve its objectives. Finally a VBE instantiation methodology is introduced as a controlled process, addressing systematically a set of steps, supported by different assisting means (e.g. methods and tools) for establishing the management functionalities and running of a VBE during its entire lifecycle, on the basis of the VBE reference model and VBE management framework proposed.