Method for producing mesenchymal stem cells with enhanced proliferative potential

Abstract

Method for producing mesenchymal stem cells with enhanced proliferative potential, in which mesenchymal stem cells are isolated from the connective tissue of an umbilical cord of a newborn by the explant method, using an umbilical cord obtained during normal childbirth after cutting or thawed after cryopreservation, from which the blood vessels have been removed. The explants are incubated in a CO₂ incubator under hypoxic conditions, with the hypoxic mixture consisting of 5% CO₂, 5% oxygen, and 90% nitrogen, and are maintained throughout the entire cell culture cycle. The culture medium is changed twice a week. After the release of zero-passage cells that formed clones, the explants are removed from the flask. When the clones reach ±80-90% confluence, the cells are transferred to new flasks using standard procedures, and the first passage is cultured under hypoxic conditions as well. Similarly, under hypoxic conditions, mesenchymal stem cells with enhanced proliferative potential are obtained in the second passage and used for transplantation in experiments involving second-passage mesenchymal stem cells for multiple sclerosis and cognitive impairments.