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.