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. 2008 Oct;58(2):69-75.
doi: 10.1007/s10616-008-9174-8. Epub 2008 Nov 7.

Temperature and calcium ions affect aggregation of mesenchymal stem cells in phosphate buffered saline

Boon C Heng  1 , Catherine M Cowan, Shubhayu Basu
Affiliations

Temperature and calcium ions affect aggregation of mesenchymal stem cells in phosphate buffered saline

Boon C Heng et al. Cytotechnology. 2008 Oct.

Abstract

Bone marrow-derived mesenchymal stem cells (MSC) are being extensively studied as potential therapeutic agents for various diseases and have demonstrated tremendous promise to date. To reduce immunological and inflammatory reaction upon delivery of MSC in situ, the cells are often suspended in protein-free and nutrient-poor buffered saline solution at high titers and kept on ice (0 degrees C) until completion of the transplantation procedure. This study investigated the effects of suspending MSC (5 x 10(6) cells/mL) in phosphate buffered saline (PBS) with and without calcium, over a time course of 90 and 180 min, at temperatures of 0 and 37 degrees C. The results at 0 degrees C showed a small but significant decrease in cell viability within calcium-free PBS after 180 min, whereas no significant changes in cell viability were observed with PBS containing calcium. Additionally, it was observed that significant aggregation of MSC into cellular clumps occurred when incubated in PBS at 0 degrees C, with a higher degree of aggregation occurring under calcium-free conditions. By contrast at 37 degrees C, there was a more pronounced decrease in cell viability after 90 and 180 min, but lesser aggregation of MSC both in the presence and absence of calcium. The aggregation of MSC into cellular clumps could pose an embolic hazard if delivered into the arterial vasculature in cardiac applications, can clog-up injection or infusion catheters utilized for cell delivery during surgery, and can also possibly reduce the overall efficacy of transplantation therapy.

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Figures

Fig. 1
Fig. 1
MSC incubated at 0 °C in a PBS without calcium after 0 min, b PBS without calcium after 90 min, c PBS without calcium after 180 min, d PBS with calcium after 0 min, e PBS with calcium after 90 min, f PBS with calcium after 180 min
Fig. 2
Fig. 2
MSC incubated at 0 °C were analyzed for cell viability and titer with the Vi-CELL® XR analyzer at the 0, 90 and 180 min time points. a cell titer after incubation for 90 and 180 min, expressed as a percentage of the initial reading at zero time point, b cell viability at the 0, 90 and 180 min time points
Fig. 3
Fig. 3
MSC incubated at 37 °C in a PBS without calcium after 0 min, b PBS without calcium after 90 min, c PBS without calcium after 180 min, d PBS with calcium after 0 min, e PBS with calcium after 90 min, f PBS with calcium after 180 min
Fig. 4
Fig. 4
MSC incubated at 37 °C were analyzed for cell viability and titer with the Vi-CELL® XR analyzer at the 0, 90 and 180 min time points. a cell titer after incubation for 90 and 180 min, expressed as a percentage of the initial reading at zero time point, b cell viability at the 0, 90 and 180 min time points
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References

    1. Abdel-Latif A, Bolli R, Tleyjeh IM, Montori VM, Perin EC, Hornung CA, Zuba-Surma EK, Al-Mallah M, Dawn B (2007) Adult bone marrow-derived cells for cardiac repair: a systematic review and meta-analysis. Arch Intern Med 167(10):989–997. doi:10.1001/archinte.167.10.989 - DOI - PubMed
    1. Bo T, Pawliszyn J (2006) Role of calcium binding in protein structural changes and phospholipid–protein interactions studied by capillary isoelectric focusing with whole column imaging detection. Anal Chim Acta 559:1–8. doi:10.1016/j.aca.2005.11.047 - DOI - PubMed
    1. Dezawa M (2008) Systematic neuronal and muscle induction systems in bone marrow stromal cells: the potential for tissue reconstruction in neurodegenerative and muscle degenerative diseases. Med Mol Morphol 41(1):14–19. doi:10.1007/s00795-007-0389-0 - DOI - PubMed
    1. Dynlacht JR, Fox MH (1992) Heat-induced changes in the membrane fluidity of Chinese hamster ovary cells measured by flow cytometry. Radiat Res 130(1):48–54. doi:10.2307/3578478 - DOI - PubMed
    1. Grauss RW, Winter EM, van Tuyn J, Pijnappels DA, Steijn RV, Hogers B, van der Geest RJ, de Vries AA, Steendijk P, van der Laarse A, Gittenberger-de Groot AC, Schalij MJ, Atsma DE (2007) Mesenchymal stem cells from ischemic heart disease patients improve left ventricular function after acute myocardial infarction. Am J Physiol Heart Circ Physiol 293(4):H2438–H2447. doi:10.1152/ajpheart.00365.2007 - DOI - PubMed