Threshold voltage shift under applied gate voltage is a key factor characterizing stability of organic thin-film transistors (OTFTs), while the physical mechanism is still controversial. In this study, we systematically examined the initial growth of pentacene polycrystalline films under different growth rates. Bias stress performance of the fabricated pentacene-based OTFTs was found to be highly related to the initial gain size of the pentacene films. Larger grain size at the first deposition layer led to smaller threshold voltage shift. The quantitative correlation can be described by a two-dimensional microscopic mobility model relating to the grains and grain boundaries.