Human milk can be an important source for obtaining potential probiotics strains for newborns in order to establish the beneficial gut microbial community and development of immune system. The aim of the study was to explore potential human breast milk probiotics and to carry out their in vitro biosafety assessment. The study obtained three isolates namely, SP₁B, B₂Enr and SP₁ which showed potential probiotic activities compared to standard probiotic Lactobacillus plantarum. In addition, these isolates were found to be safe through various in vitro biosafety aspects. The molecular identification by16srDNA sequencing revealed that SP₁B and B₂Enr belong to Bacillus cereus (MK210172) and Staphylococcus epidermidis (MK210234), respectively. For the first time, the study suggests that these bacterial strains may come in the category of probiotics and can be considered further after in vivo biosafety assessments.