Abstract
Aimed at providing a contribution to the optimization of cryopreservation processes, the present work focuses on the osmotic behavior of human mesenchymal stem cells (hMSCs). Once isolated from the umbilical cord blood (UCB) of three different donors, hMSCs were characterized in terms of size distribution and their osmotic properties suitably evaluated through the exposure to hypertonic and isotonic aqueous solutions at three different temperatures. More specifically, inactive cell volume and cell permeability to water and di-methyl sulfoxide (DMSO) were measured, being cell size determined using impedance measurements under both equilibrium and dynamic conditions. Experimental findings indicate that positive cell volume excursions are limited by the apparent increase of inactive volume, which occurs during both the shrink-swell process following DMSO addition and the subsequent restoration of isotonic conditions in the presence of hypertonic solutions of impermeant or permeant solutes. Based on this evidence, hMSCs must be regarded as imperfect osmometers, and their osmotic behavior described within a scenario no longer compatible with the simple two-parameter model usually utilized in the literature. In this respect, the activation of mechano-sensitive ion-channels seemingly represents a reasonable hypothesis for rationalizing the observed osmotic behavior of hMSCs from UCB.
Highlights
The progressive ageing of population in western and industrialised countries represents one of currently emerging demographic trends with straightforward implications for well-being and welfare policies
Dynamic runs were performed at 17, 27 and 37 ̊C with cells injected into a Osmosis of human mesenchymal stem cells hypertonic solution with 600 or 2000 mOsm/L osmolarity depending on the use of non-permeant sucrose and permeant di-methyl sulfoxide (DMSO) respectively
The 20% success rate is consistent with literature data [32], and can be expected to depend mostly on time elapsed from birth to isolation and size of umbilical cord blood (UCB) units [2,13]
Summary
The progressive ageing of population in western and industrialised countries represents one of currently emerging demographic trends with straightforward implications for well-being and welfare policies. Dynamic runs were performed at 17, 27 and 37 ̊C with cells injected into a Osmosis of human mesenchymal stem cells hypertonic solution with 600 or 2000 mOsm/L osmolarity depending on the use of non-permeant sucrose and permeant DMSO respectively.
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