The establishment of gene banks using cryopreservation to secure the genetic diversity of farm breeds have been widely assessed. France, the Netherlands and the USA were among the first countries to organize national cryobanks and these banks are now 10 to 20 years old. All three countries have started Holstein-Friesian (HF) collections to conserve as much genetic diversity as possible for this globally important breed. In order better understand the diversity captured in these collections, the genetic variability of HF collections within and between countries was assessed, and genetic variability of germplasm collections were compared with active bulls in each country. The overall aim of the project was to determine the breed’s security and to guide future collection activities.