Figure 6

MACS+ cells’ role in cultured ISMC Mix with rhythmic contractions in vitro. (a–c) 100 k MACS0 (passaged ISMC Mix: mixture of MACS+ cells, SMCs and neuronal cells) or MACS− cells (ISMC Mix without MACS+ cells: mostly SMCs and neuronal cells) were cultured on STO cells for 3 weeks. Cells were seeded and cultured in FBS medium for the first 4 days before changing to F12 medium. (d–f) 100 k MACS− were seeded on STO cells for 5 weeks, with (MACS−/+) or without (MACS−) addition of 60 k MACS+ cells on day 5. Cells were cultured in FBS medium for the first 7 days before changing to F12 medium. (a,d) Confocal images of ICC markers, Kit (red), Ano1 (green), co-localization (yellow). Scale bar, 100 µm. (b,e) Frequency of cultured cells motility due to spontaneous contraction (b: wk2 n = 2, wk3 n = 4; (e) wk5 n = 4). (c,f) Cultured cells were analyzed for mRNA expression (Kit, Myh11, Tubb3) at week 2 (c) or week 5 (f) (n = 4). *Samples were normalized to de-epithelialized intestine. Error bars, s.d. ***P < 0.0001, **P < 0.01, *P < 0.05.