Creation and Characterization of Human Mesenchymal Stromal Cell Cultures with Prolonged Proliferative Potential for Regenerative Medicine
摘要
Objectives: The study of molecular mechanisms of regeneration requires convenient models for in vitro and in vivo studies. In vitro cell cultures can fulfill such a function. However, their rapid aging and loss of initial specific properties in culture is a significant limitation of their use. Telomerase expression can help to overcome these limitations: it can prolong proliferative activity and stabilize the initial properties of a primary cell culture. Methods: Here, we created and studied the properties of human adipose tissue multipotent mesenchymal stromal cell (MSC) cultures that overexpress the catalytic subunit of human telomerase (hTERT). Results: We found that these MSC cultures were able to proliferate for 38–63 population doublings (PD), maintained sensitivity to noradrenaline, serotonin, glutamate, γ-aminobutyric acid, parathyroid hormone, angiotensin II, and histamine until at least 26 PD, retained an MSC-specific immunophenotype until at least 36 PD, and preserved the ability to undergo adipogenic, osteogenic, and chondrogenic differentiation until at least 39 PD. Moreover, overexpression of hTERT in MSC cultures stabilized the qualitative and quantitative composition of their secretome at long-term passaging (at least up to 30 PD). Conclusions: The obtained results allow us to consider telomerase hyperexpression as a promising approach to obtaining MSC cultures with prolonged proliferative activity, which can be used as a stable and convenient object for fundamental and applied studies in the field of regenerative medicine.