Figure 1
From: Bioengineering functional smooth muscle with spontaneous rhythmic contraction in vitro
Maintenance of MACS+ cells cultured on STO cells. (a) Confocal images of ICC markers, Kit (red), Ano1 (green), and co-localization (yellow). 60 k MACS+ and 15 k MACS+ cells were cultured for 7 days and 14 days respectively. Scale bar, 100 µm. (b) MACS+ cells were analyzed for Kit mRNA expression (b–d: day 1, 4, 7 n = 4; day 14 n = 2). (c,d) GFP + MACS+ cells were seeded on STO cells, MEF, and Ge, cultured, and analyzed for mRNA expression of Kit (c) and gfp DNA (d). STO cells and MEF do not express Kit/gfp. (e) mRNA expression of Kit for MACS+ cells cultured on STO cells, MEF and Ge for 7 days. (f) mRNA expression of Kit for MACS+ cells cultured on different STO seeding densities for 7 days, where 100% is 100 k (100%, n = 5; 50%, n = 4; 25%, n = 2). (g) mRNA expression of Kit for MACS+ cells cultured on Ge for 7 days in media supplemented with 25, 50, or 100 ng/ml of soluble scf (n = 2). STO and Ge were controls (n = 5). (h) mRNA expression of Kit for 60 k or 15 k MACS+ cells cultured on Ge for 7 days supplemented with conditioned media from STO (Ge-CM), where Ge was the control (n = 4). STO = Mouse Embryonic Fibroblfast (Sandos Inbred Mouse, SIM). MEF = Mouse Embryonic Fibroblast (C57BL/6). Ge = gelatin coating. Feeder cells (STO, MEF) were mitomycin C treated. *Samples were normalized to de-epithelialized intestine. Error bars, s.d. ***P < 0.0001, *P < 0.05.
