Journal: Biomedical Materials (Bristol, England)
Article Title: Improvement of cellular pattern organization and clarity through centrifugal force
doi: 10.1088/1748-605X/ada508
Figure Lengend Snippet: Long-Term Functionality through Tube Formation Assays. (A) Representative images of MPCs seeded on collagen 1 in differentiation media directly from culture (negative control, left) and of MPCs seeded on collagen 1 fully conjugated with biotin-streptavidin and centrifuged 2 × 1 min at 43.3 RCF then placed in differentiation media (patterning + centrifugation, right). Samples were fixed on Day 8 and stained with anti-myosin (green) and DAPI (blue), then imaged using fluorescent microscopy at 10x magnification to visualize tube formation. (B) Average myotube length and number of nuclei per myotube for each condition described in (A). Data is shown as mean ± standard deviation of n = 3 independent samples for each condition. Significance testing compared the negative control to patterning + centrifugation using a two-group independent t-test (* p < 0.05, ** p < 0.01, *** p < 0.001). The absence of * is considered a nonsignificant p-value comparison. (C) Representative images of HUVEC cells seeded on Matrigel in EGM TM −2 BulletKit TM media directly from culture (negative control, left) and of HUVECs seeded on Matrigel fully conjugated with biotin-streptavidin and centrifuged 2 × 1 min at 43.3 RCF then placed in EGM TM −2 media (patterning + centrifugation, right). Samples were left in media overnight then stained with Calcein AM and imaged using fluorescent microscopy at 10x magnification to visualize tube formation. (D) Quantitative analysis of HUVEC tube formation including the number of nodes, junctions, segments, and branches formed from samples of each condition in (C). Data is shown as mean ± standard deviation of n = 3 independent samples for each condition. Significance testing compared the negative control in media, the negative control in PBS, and the patterning + centrifugation for each metric using a two-group independent t-test (* p < 0.05, ** p < 0.01, *** p < 0.001). The absence of * is considered a nonsignificant p-value comparison. All scale bars represent 200 μm.
Article Snippet: HUVECs were cultured in EGM TM −2 endothelial growth media BulletKit TM (Lonza, Basel, Switzerland) supplemented with 1% penicillin-streptomycin and 1% antibiotic-antimyocotic.
Techniques: Negative Control, Centrifugation, Staining, Microscopy, Standard Deviation, Comparison
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