IntroductionPasteurized human donor milk (DM) is frequently used for feeding preterm newborns and extrauterine growth‐restricted (EUGR) infants. Most human milk banks performed a pasteurization of DM using the standard method of Holder pasteurization (HoP) which consists of heating milk at 62.5°C for 30 min. High hydrostatic pressure (HHP) processing was proposed to be an innovative nonthermal method to pasteurize DM. However, the effect of different modes of DM pasteurization on body growth, intestinal maturation, and microbiota has never been investigated in vivo during the lactation.ObjectivesWe aimed to study these effects in postnatally growth‐restricted (PNGR) mice pups daily supplemented with HoP‐DM or HHP‐DM.MethodsPNGR was induced by increasing the number of pups per litter (15 pups/mother) at postnatal Day 4 (PND4). From PND8 to PND20, mice pups were supplemented with HoP‐DM or HHP‐DM. At PND21, the intestinal permeability was measured in vivo, the intestinal mucosal histology, gut microbiota, and short‐chain fatty acids (SCFAs) level were analyzed.ResultsHHP‐DM pups displayed a significantly higher body weight gain than HoP‐DM pups during lactation. At PND21, these two types of human milk supplementations did not differentially alter intestinal morphology and permeability, the gene‐expression level of several mucosal intestinal markers, gut microbiota, and the caecal SCFAs level.ConclusionOur data suggest that HHP could be an attractive alternative to HoP and that HHP‐DM may ensure a better body growth of preterm and/or EUGR infants.