Heart rate variability (HRV) analysis is an established method to characterize the autonomic regulation and is based mostly on 24h Holter recordings. The importance of short-term HRV (less than 30 min) for various applications is growing consistently. Major reasons for this are the suitability for ambulatory care and patient monitoring and the ability to provide an almost immediate test result. So far, there have been only a few studies that provided statistically relevant reference values for short-term HRV. In our study, 5 min short-term HRV indices were determined from 1906 healthy subjects. From these records, linear and nonlinear indices were extracted. To determine general age-related influences, HRV indices were compared from subjects aged 25-49 years with subjects aged 50-74 years. In a second approach, we examined the development of HRV indices by age in terms of age decades (25-34, 35-44, 45-54, 55-64 and 65-74 years). Our results showed significant variations of HRV indices by age in almost all domains. While marked dynamics in terms of parameter change (variability reduction) were observed in the first age decades, in particular the last two age decades showed certain constancy with respect to the HRV indices examined.