Startup, or start-up, is a business entity, typically described as a newly established company, which is based on an innovative business concept using advanced technologies and is rapidly evolving. Start-up has large potential for economic growth. These firms are often funded by their business founders and venture scapitalists. Due to high costs or limited revenues, a huge number of these small companies are not able to be maintained at a certain rate or level in the long term without additional funding by venture funds, business angel or private or public sector. Starup companies cooperate in networks where they use access to supply of money, materials, staff, and other assets that can be drawn on by a person or organization in order to function effectively through different sort of stakeholders and key actors. It seems not clear to understand all particular issue and entities which these companies should concentrate their networking endeavour on, especially in correlation of variable of inputs is important to describe and see the connotation how the regional innovation strategy influences survival at their early stage. The article analyses a unique sample of 30 Czech start-up companies established between 2010 and 2020. The results show that the survival of a start-up company depends on three main part of network of actors: investors, external entrepreneurs, entrepreneur in residence and an influence of innovation centre (or science parks). In addition, start-ups that were raised in the region with a regional innovation strategy summarize the objectives, problem areas and priorities that will be required to deliver regional development policy, including business growth and innovation and have shown a greater likelihood of survival.