Background Autologous peripheral blood stem-cell collection (PBSCC) in children has become an integral part of contemporary treatment protocols, but the procedure is often complicated due to technical issues related to vascular access. Central line placement is often implemented to surmount this problem, but is associated with complications such as bleeding, thrombosis and pneumothorax. As an alternative we have introduced the use of radial arterial lines for PBSCC in children. Procedure Data from autologous stem cell collections performed from October 2002 to December 2011 using a radial arterial line were collected. Results A total of 372 PBSCC procedures were performed during the study period; an arterial line was used in 311 PBSCC's in 208 children. The average patient age and weight were 7.9 years (SD 5.4) and 28.3kg (SD 20.4), respectively. The smallest patient was 9 months old and weighed 7kg. The mean total volume processed was 8,593cm3 (SD 4,854), and the mean number of blood volumes processed was 4.3. Mean collection time for a single blood volume was 55minutes (SD 15.5). The mean number of CD34+ cells collected per donation was 5.8x106/kg. Ninety-seven patients (46%) required more than one collection to meet the requested CD34+ cell target. No serious adverse effects associated with vascular access occurred in this cohort. Conclusion Percutaneous placement of radial artery catheters can be rapidly and safely performed in very small infants and in children with difficult venous access. This technique provides a reliable platform for efficient PBSCC. Pediatr Blood Cancer 2013; 60: 946948. (c) 2012 Wiley Periodicals, Inc.