Recovery of Human Mesenchymal Stem Cells Following Dehydration and Rehydration

• Published in: Cryobiology 2001-09, Vol.43 (2), p.182-187
• Main Authors: Gordon, Stephen L., Oppenheimer, Stephanie R., Mackay, Alastair M., Brunnabend, Jason, Puhlev, Iskren, Levine, Fred
• Format: Article
• Language: English
• Subjects: Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Antigens, Surface - metabolism; Applied cell therapy and gene therapy; Biological and medical sciences; Cell Count; Cell Differentiation; Cell physiology; Cell Survival; dehydration; Desiccation - methods; Effects of physical and chemical agents; Flow Cytometry; Fundamental and applied biological sciences. Psychology; glycerol; human mesenchymal stem cells; Humans; In Vitro Techniques; Medical sciences; Mesoderm - cytology; Mesoderm - immunology; Molecular and cellular biology; Reverse Transcriptase Polymerase Chain Reaction; Stem Cells - cytology; Stem Cells - immunology; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; trehalose; vacuum
• ISSN: 0011-2240; 1090-2392
• DOI: 10.1006/cryo.2001.2361

As cell therapies advance from research laboratories to clinical application, there is the need to transport cells and tissues across long distances while maintaining cell viability and function. Currently cells are successfully stored and shipped under liquid nitrogen vapor. The ability to store these cells in the desiccated state at ambient temperature would provide tremendous economic and practical advantage. Human mesenchymal stem cells (hMSCs) have broad potential uses in tissue engineering and regeneration since they can differentiate along multiple lineages and support hematopoeisis. The current research applied recent technological advances in the dehydration and storage of human fibroblasts to hMSCs. Three conditions were tested: air-dried, air-dried and stored under vacuum (vacuum only), and incubated with 50 mM trehalose + 3% glycerol and then air-dried and stored under vacuum (vacuum + trehalose). Plates containing dehydrated hMSCs were shipped from San Diego to Baltimore overnight in separate FedEx cardboard boxes. The hMSCs were rehydrated with 3 ml of hMSC medium and were able to regain their spindle-shaped morphology and adhesive capability. In addition, they maintained high viability and proliferation capacity. Rehydrated and passaged cells continued to express the characteristic hMSC surface antigen panel. Additionally, cells showed constitutive levels of mRNA for a stromal factor and, when exposed to reagents known to induce differentiation, demonstrated upregulation of two tissue-specific messages indicative of differentiation potential for fat and bone. While our preliminary findings are encouraging, we still need to address consistency and duration of storage by considering factors such as cell water content, oxygen concentration, and the presence of free radicals.