Morphological and functional characteristics of cryopreserved multipotent mesenchymal stromal cells from bone marrow, adipose tissue and tendons

Abstract

The aim of study was to comparatively evaluate the morphological and functional properties of cryopreserved multipotent mesenchymal stromal cells (MMSCs) from bone marrow, fat and tendon.Materials and methods. MMSC cultures obtained from rat bone marrow, fat and tendon. The cells was cryopreserved under protection of 10 % DMSO and 20% FBS with cooling rate of 1 deg/min down to -80°C followed by plunging into liquid nitrogen. In the studied cultures the membrane integrity, immunophenotype, ability to colony formation, proliferative characteristics (MTT-test), directed differentiation and type I collagen synthesis were evaluated.Results. Investigated cryopreserved cell culture derived from bone marrow, tendon and adipose tissue had high membrane integrity indicators, colony formation and proliferation as well as the ability to directional adipogenic and chondrogenic differentiation. The analysis of immunophenotype showed that the tested cryopreserved MMSCs culture characterized by high levels of expression (≥90 %) of CD44, CD90, CD105, CD73 and low expression (≤1 %) of hematopoietic marker CD45. Cryopreserved bone marrow MMSCs were characterized by a high content of cells that synthesized type I collagen as compared to cultures which were derived from fat and tendon.Conclusions. Cell cultures derived from all studied sources have immunophenotype of precursor cells of mesenchymal origin. The MMSC of tendon tissue characterized by a greater capacity for colony formation and proliferation, and lower capacity for directed adipogenic differentiation, than MSCs from bone marrow and adipose tissue.