Biopreservation and Biobanking - Ziath - 16

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ANGEL ET AL.
FIG. 3. PBMC viability after thawing (A) and after overnight culture (B). Figures show the PBMC viability after 0-350
temperature cycles. PBMC viability determined directly after thawing and after overnight culture. Figures show
mean - standard deviation. *Statistical difference to 0 cycles with p< 0.05. PBMC, peripheral blood mononuclear cell.
medium responder, and one donor was classified as high
responder (Table 1). After stimulation with CMV, none of
four donors were classified negative, two donors as low
responders, and two donors as high responders (Table 2).
Donor 1 showed a high response on CEF as well as on
CMV stimulation. Except for 350 cycles, the T cell response
decreased with increasing numbers of temperature cycles
(Tables 1 and 2). For Donor 2, a low responder, a decrease
with increasing numbers of temperature cycles after both
CEF and CMV stimulation was also observed. Donors 3,
apart from 350 cycles, and 4 showed similar behavior (Tables
1, 2). After stimulation with CEF, the decrease of the
high responder cells became significant after 250 and 300
cycles and medium responder's cells after 200 cycles. The
low responder cells showed no significant degradation.
After stimulation with CMV peptide pool, the decrease in
high responder cells of Donor 1 was significant after 200,
250, and 300 temperature cycles and for Donor 3 after 100
cycles. The low responder cells showed no statistically
significant decrease.
To present the decrease in a clearer manner, we pooled
and normalized the data of all donors. A linear regression
model was used to calculate the relative reduction per 50
cycles in percent (%). After stimulation with either CEF or
CMV peptides, the immune response decreased with increasing
numbers of temperature fluctuations. After 50 cycles,
the T cell response decreased about -11.39% - 9.52%
after CEF stimulation and about -7.38% - 8.73% (Fig. 5A).
The decrease reached a maximum of -27.70% - 13.80%
after CEF stimulation (Fig. 5A) and -22.52% - 11.44% after
CMV stimulation after 350 temperature cycles (Fig. 5B).
The pooled data of each 50th cycle group show no significant
differences (N= 12, one-way ANOVA Bonferroni
with p< 0.05, R2 [CEF] = 0.50355, R2 [CMV]= 0.36517).
FIG. 4. PBMC recovery after thawing (A) and after overnight culture (B). Figures show the PBMC recovery after 0-350
temperature cycles. PBMC recovery determined directly after thawing and after overnight culture. Figures show
mean - standard deviation. *Statistical difference to 0 cycles with p< 0.05.

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