Figure 3
From: Bioengineering functional smooth muscle with spontaneous rhythmic contraction in vitro
Application of MACS+ cells cultured on STO cells. (a) Confocal images of ICC markers, Kit (red), Ano1 (green), and co-localization (yellow). 15 k MACS+ cells were cultured on STO-seeded ePCL scaffold for 14 days. Scale bar, 100 µm. (b,c) Quantification of Kit alignment expressed by 15 k MACS+ cells cultured on STO-seeded ePCL and glass for 14 days. (b) Immunofluorescence of ICC markers, Kit (red) and Ano1 (green) and with co-localization (yellow). Scale bar, 200 µm. (c) Coherency analysis of Kit expression of MACS+ cells cultured on ePCL(aligned) and glass (random), where higher coherency means greater cell alignment (n = 5). (d,e) GFP + MACS+ cells were cultured on STO cells for 4 days and purified using MACS before colonoscopic injection into the submucosa of the rectum of C57BL/6 mice. A permanent carbon ink was mixed into the cell suspension to mark injection sites. The injected rectums were retrieved after 7 days, and were immunostained for Ano1 (d, red), α-SMA (e, red) and GFP (d,e, green). GFP co-localized with Ano1 (d, yellow) but not with α-SMA (e). Merged immunofluorescence images (left) were further merged with phase contrast images to show the area of injection indicated by black ink (right). Scale bar, 100 µm. Insets, higher-magnification images of boxed regions. Scale bar, 10 µm. Error bars, s.d. ***P < 0.0001.
