PA5 hOMCs phenotype and promotion of neuron outgrowth after
growth on Plastic and Plastic Plus microcarrier suspension culture
RT-qPCR results (Figure 4 A and B), show the upregulation of several
markers for PA5 hOMCs grown on Plastic Plus compared to cells at day 0.
Fibronectin (3.5-fold), β-III tubulin (2.3-fold), KDR1 (2.9-fold) and
Sox9 (2.9-fold) are upregulated for PA5 hOMCs grown on Plastic Plus
microcarriers compared to PA5 hOMCs grown on Plastic microcarriers by
day 7. Most importantly, p75NTR, a neurotrophic marker
expressed by OECs is expressed 2.3-fold more on day 7 by PA5 hOMCs grown
on Plastic Plus microcarriers compared with cells on day 0, while PA5
hOMCS grown on Plastic microcarriers maintained the same level of
expression of this marker (1.0-fold) (p<0.05). Furthermore, a
co-culture assay with NG108-15 cells was used as an assay indicative of
the capacity of PA5 hOMCs to promote neurite outgrowth, using F7 Schwan
cells as positive control and NG108-15 cell only as a negative control.
Representative images used to quantify neurite outgrowth are shown in
Figure 4 C. The average neurite length was 36±1.3 µm for Plastic Plus
microcarriers, significantly higher (p<0.05) than the average
neurite length obtained for Plastic (29.1±1.5 µm) microcarriers and the
positive control (27.9±0.1 µm) (Figure 4 D). The maximum neurite length
obtained for Plastic Plus was 96±7.0 µm, significantly higher
(p<0.05) than the maximum neurite length obtained for Plastic
microcarriers (81.8±3.2 µm) and the positive control (57.9±3.3 µm)
(Figure 4 E). The average neurite per neuron was of 0.38±0.05,
significantly higher (p<0.05) than the one obtained for cells
grown on Plastic microcarriers (0.23±0.02) and the positive control
(0.22±0.04) (Figure 4 F).