The purpose of the research was to study the behavior of several Acer varieties: Acer palmatum 'Dissectum Atropurpureum' and Acer platanoides 'Crimson King', 'Drummondii' and 'Globosum', during the first subculture of the in vitro multiplication phase. There were used explants from the initiation phase and passed on the multiplication nutrient media. The nutrient media tested were MS, DKW and WPM and each had a different content of growth hormones. In order to determine the multiplication rate, was tested the influence of zeatin and thidiazuron, in concentration of 1.5 mg/l, on a constant level of indole-butyric acid (0.5 mg/l). In the growing chamber were a constant temperature and a photoperiod of 14 hours light/24 hours, at a luminosity of 2500 lux. From the observations and registered data was determined the multiplication rate (microshoots/explant) for the four genotypes.