Iniciation and cultivation of multipotent mesenchimal human umbilical stroma cells in a laboratory experiment

BACKGROUND: Wharton’s jelly of the human umbilical cord is one of the sources of multipotent mesenchymal stromal cells. The cell population obtained from the postpartum biomaterial is characterized by high proliferative and regenerative properties. Isolation of a culture of multipotent mesenchymal stromal cells from the umbilical cord does not pose a threat to the health and life of the donor.

AIM: Optimization of the technique for isolating a reproducible population of multipotent mesenchymal stromal cells from Wharton’s jelly is an urgent task in biomedicine, which can accelerate the process of obtaining donor cells for cell therapy and tissue engineering.

MATERIALS AND METHODS: In the study, the main techniques and methods for isolating the culture of umbilical cord stroma cells were tested, the cultivation process was optimized to increase its efficiency and reduce the time of growth of cell biomass. The effect of the components of the nutrient medium on the cells obtained from Wharton’s jelly of the human umbilical cord was studied. Currently, there is no universal composition of the growth medium; in various studies, nutrient media from different manufacturers are used, which differ in composition. The most discussed issue is the selection of serum, which is part of the nutrient medium.

RESULTS: In the work, a comparative evaluation of five different sera was carried out. It has been shown that the most stable physiological parameters are observed in cell suspension samples with the addition of FBS (SKPK, Russia) and FBS (Capricorn, USA) sera. A study of the effect of hypoxia on cell culture in combination with the most effective sera showed that hypoxic stress acts as an activator of primary cell proliferation. The assessment of the effect of serum and hypoxia on cell culture was carried out visually using microscopy, assessment of changes in cell morphology during cultivation, and the results of testing the action of sera by the intensity of respiration of free and immobilized cells under the action of inhibitors.

CONCLUSION: As a result of the experiments, the influence of the type of serum on the initiation of cell expansion from primary explants and further cell proliferation in vitro was established. Hypoxia during exposure of primary explants enhances the expansion of cells from tissue fragments of Wharton’s jelly tissue.