Page 83 - Human Umbilical Cord Mesenchymal Stem Cells

Umbilical Cord MSC to Support Cell Transplantation 1483

1035
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776 0.9% Blank
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776 GFP DNA (15µg)
Counts 517 30.9% 300mV 150µF
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776 68.0% GFP RNA (5µg)
Counts 517 300mV 150µF
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776 81.2% GFP RNA (5µg)
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Counts 517 89.8% GFP RNA (5µg)
300mV 300µF
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Figure 3. Transfection of UC-MSCs with the GFP reporter gene. Adherent cells were transfected using the BioRad
electroporator with either cDNA or mRNA at different voltages and capacitance, as indicated. mRNA showed higher trans-
fection efficiency than cDNA, with expression of the GFP gene maintained for several days.

plasma avoids exposure of UC-MSCs to either animal This facilitates the use of autologous UC-MSCs for
serum or allogeneic HS. Both serum preparations are any medical indication (beyond transplantation) at
known to alter the gene profile and antigen expres- a later point. Our results also confirm that large num-
sion of MSCs [27,28]. In addition, FBS or allogeneic bers of UC-MSCs can be expanded in culture from
HS potentially could be contaminated with viruses or a small piece of cord that has been finely minced
prions, and preserving UC tissue in autologous cord and that the microparticles can be cryopreserved on
plasma avoids those risks. Mincing and freezing the arrival in the laboratory. When preparing the cord,
cord tissue in autologous cord serum is of practical special care was taken not to collect the perivascular
importance from a UCB banking perspective, because cells in the cord (pericytes), which have different
this method avoids previous culture expansion of characteristics despite being MSCs [9]. Our cells
UC-MSCs. It allows for freezing of the UC-MSCs were consistently negative for the 3G5 surface anti-
at the same time as the UCB arrives, and both cell gen, suggesting that our preparations were free of
types can be stored in the same dual-chamber bag. pericytes.