Proliferative scarring is a human disease with neither available effective treatment nor relevant animal model. One of the hypotheses for scar formation involves deregulation of fibroblast signaling and delayed apoptosis. Here, we introduce a new chemical-free method for fibroblast density control in culture by intermittently delivered pulsed electric fields (IDPEF), which cause irreversible damage to cell membranes. Using 5–100 pulses with electric field strength of 150 V mm−1, pulse duration 70 µs, and frequency of 1 Hz, we investigated the effects of pulsed electric field application on growth, death, and regeneration of normal human dermal fibroblasts in culture. We found that the fraction of fibroblasts that survive depends on the number of pulses applied and follows a Weibull distribution. We have successfully developed an IDPEF protocol that controls fibroblasts density in culture. Specifically, through application of IDPEF every 72 h for 12 days, we maintain a normal human dermal fibroblast density in the 3.1 ± 0.2 × 105 to 1.4 ± 0.2 × 105 cell mL−1 range. Our results suggest that IDPEFs may prove useful as a non-chemical method for fibroblast density control in human wound healing. Biotechnol. Bioeng. © 2013 Wiley Periodicals, Inc.