Figure Legends
Figure 1 - PA5 hOMCs were grown on microcarriers in 96-well plates for 7
days on 10 different microcarriers. (A) Growth curve showing viable
cells/well obtained through measurements of metabolite activity using
the CCK-8 viability reagent. (B) Total viable cell/well achieved for
each microcarrier on day 1. (C) Total viable cell/well achieved for each
microcarrier on day 7. Data represented as mean ± SEM, n=3. Significant
differences were noted with (*) for p<0.05.
Figure 2 – PA5 hOMCs were grown on Plastic L, Synthemax II LC,
Collagen, Plastic, PronectinF and Plastic Plus microcarriers for 7 days
with 50% media change every 2 days and using an initial seeding density
of 6000 cells/cm2. Viable cell concentration (A) and
cell viability (B) were measured. Metabolite analysis for glucose (C),
lactate (D), and ammonium (E) were performed. (F) Expression of
p75NTR, S100β, GFAP, β-III tubulin, nestin and
fibronectin by PA5 hOMCs grown on Plastic L, Synthemax II LC, Collagen,
Plastic, PronectinF and Plastic Plus, assessed through RT-qPCR on day 7.
PA5 hOMCs before seeding were used as a reference.Data represented as
mean ± SEM, n = 3. Significant differences were noted with (*) for
p<0.05.
Figure 3 - Comparison of PA5 hOMCs growth on Plastic and Plastic Plus
microcarriers, in spinner flasks for 7 days. (A) Viable cell
concentration (cells/mL) and cell viability was measured via
haemocytometer counts. Metabolite analysis was performed to determine
concentration of (B) glucose (mM), (C) lactate (mM) and (D) ammonium
(mM). (E) Representative images of cells on Plastic and Plastic Plus
microcarriers, for each day of sample. Hoechst was used as a nucleic
dye. Data is presented as mean ± SEM, n=3. Significant differences were
noted with (*) for p<0.05. Scale bar = 200 µm.
Figure 4 –The expression of p75NTR, GFAP, S100β
(glial markers), β-III tubulin, nestin (neural markers), fibronectin
(fibroblastic marker), CD90, CD105, CD73 (positive MSCs markers), CD34,
CD45 (negative MSCs markers), PPARg (adipogenesis marker), SPP1, Runx2
(osteogenesis markers), Sox9 (chondrogenesis marker) and VEGFR2
(endothelial cell marker). (A) Heat map for gene expression variation
relative to cells before seeding. (B) Values of expression relative to
cells before seeding. NG108-15 co-culture assay to assess the potential
for neural regeneration of PA5 hOMCs. F7 Schwann cells were used as a
positive control, and NG108-15 neurons only were used as negative
control. (C) Representative images used to quantify neurite outgrowth.
(D) Average neurite length, (E) maximum neurite length (F) average
neurite per neuron, were higher for Plastic and especially Plastic Plus
microcarriers (scale bar = 400 µm). Data represented as mean ± SEM
(n=3). Significant differences were noted with (*) for p<0.05.