The article aims to identify the development phases in the process of the regional differentiation in the Czech Republic after 1989, and examines whether this differentiation trend is currently changing. The assumption the article's hypothesis is based on is that during recent development the basic 'parameters' of the principal features of the country's regional structure have been stabilized amidst the conditions of parliamentary democracy and a market economy. Unlike the first phases of socio-economic transition, when differentiating trends prevailed, a certain degree of stabilisation can now be assumed, alongside the emergence of new trends. The development of regional differences was analysed using indicators of GDP, the unemployment rate, entrepreneurial activity, and tax revenue from self-employed physical persons. The findings show that over the course of the 1990s regional differences intensified at both the mezo-regional (regional) level and the micro-regional level. It was also confirmed that at the turn of the millennium the phase of divergent regional development ended, and since that time regional differences have hovered around the same level.