Trehalose preincubation increases mesenchymal (CD271+) stem cells post-cryopreservation viability
Abstract: Dimethyl sulfoxide
(Me2SO) is a common cryoprotective agent widely used in cell preservation
system. Me2SO is currently known to cause epigenetic changes which are critical in stem cells development and
cellular differentiation. Therefore, it is imperative to develop
cryopreservation techniques that protect cellular functions and avert Me2SO
adverse effect. Trehalose was able to protect organism in extreme condition
such as dehydration and cold. This study aimed to verify the protective effect
of trehalose preincubation procedure in cryopreservation.
Methods: The study was conducted using experimental design. Thawed
mesenchymal (CD271+) stem cells from YARSI biorepository were used for the
experiment. Trehalose preincubation was performed for 1 hour, internalized
trehalose was confirmed by FTIR-ATR measurement. Three groups consisted of (1)
cryopreserved without trehalose preincubation, (2) cryopreserved with trehalose
preincubation, and (3) did not undergo cryopreservation were evaluated after 24
hours in LN2 for viability in culture. The absorbance from each group was
measured at 450 nm. The analysis performed using paired student t test.
Results: Viability of thawed mesenchymal (CD271+) stem cells that undergo
trehalose preincubation prior cryopreservation was significantly higher
(p<0.05) compared to group without trehalose preincubation. Higher viability
observed between group with trehalose preincubation compared with controlled
group suggests protection to trypsinization. Mesenchymal (CD271+) stem cells
incubated for 1 hour in 100 mM trehalose supplemented medium results in 15% trehalose loading efficiency.
Conclusion: These findings confirm the protective effect of trehalose
preincubation in cryopreservation. Future research should be directed to
elucidate the trehalose internalization mechanism and eventually the protective
mechanism of trehalose in mammalian cell cryopreservation.
Author: Indra Kusuma, Restu S.
Hadi, Bambang Kiranadi, Arief Boediono
Journal Code: jpkedokterangg160137