Quantitative assessment of the impact of cryopreservation on human bone marrow-derived mesenchymal stem cells: up to 24 h post-thaw and beyond

• Published in: Stem cell research & therapy 2020-12, Vol.11 (1), p.540-540, Article 540
• Main Authors: Bahsoun, Soukaina, Coopman, Karen, Akam, Elizabeth C
• Format: Article
• Language: English
• Subjects: Apoptosis; Bone marrow; Cell differentiation; Cell lines; Cell therapy; Cell viability; Comparative analysis; Cryopreservation; Ethylenediaminetetraacetic acid; Flow cytometry; Freezing; Human bone marrow-derived mesenchymal stem cells; Manufacturers; Manufacturing; Mesenchymal stem cells; Metabolism; Quantitative; Stem cells; Thawing
• ISSN: 1757-6512; 1757-6512
• DOI: 10.1186/s13287-020-02054-2

The effects of cryopreservation on human bone marrow-derived mesenchymal stem cells (hBM-MSCs) are still ill-defined. In this study, a quantitative approach was adopted to measure several post-thaw cell attributes in order to provide an accurate reflection of the freezing and thawing impact. Fresh and cryopreserved passage-matched cells from three different donors were discretely analysed and compared for their viability, apoptosis level, phenotypic marker expression, metabolic activity, adhesion potential, proliferation rate, colony-forming unit ability (CFUF) and differentiation potentials. The results of this study show that cryopreservation reduces cell viability, increases apoptosis level and impairs hBM-MSC metabolic activity and adhesion potential in the first 4 h after thawing. At 24 h post-thaw, cell viability recovered, and apoptosis level dropped but metabolic activity and adhesion potential remained lower than fresh cells. This suggests that a 24-h period is not enough for a full recovery. Beyond 24 h post-thaw, the observed effects are variable for the three cell lines. While no difference is observed in the pre- and post-cryopreservation proliferation rate, cryopreservation reduced the CFUF ability of two of the cell lines and variably affected the adipogenic and osteogenic differentiation potentials of the three cell lines. The data collected in this study clearly show that fresh and cryopreserved hBM-MSCs are different, and these differences will inevitably introduce variabilities to the product and process development and subsequently imply financial losses. In order to avoid product divergence pre- and post-cryopreservation, effective strategies to mitigate freezing effects must be developed and implemented.