The probiotic and technological potentials of lactic acid bacteria originating from human milk are becoming a remarkable research area. In the present study, Limosilactobacillus vaginalis MA-10 isolated from human milk was investigated in vitro for its probiotic and technological aspects. According to the results obtained in the study, MA-10 strain exhibited non-haemolytic activity and various degrees of sensitivity to most of the tested antibiotics. The strain showed good resistance to the gastrointestinal system and maintained its viability under these conditions. Its antimicrobial activity against human or clinical bacterial and fungal microorganisms and fish bacteria was determined in the range of 2.38–11.22 mm. The MA-10 strain was able to assimilate cholesterol ranging from 31.42 to 82.30%. The strain showed 9.34% ferrous-ion chelating and 32% DPPH free radical scavenging activities. These initial results from the present study confirm that L. vaginalis MA-10 may be a new source with appropriated probiotic and technological traits for various industries, and further in vivo assays.Due to the limited number of studies on L. vaginalis strains originated from human breast milk in the literature, the data obtained in this study are thought to be important for revealing the basic probiotic properties of the strain.