Figure 7.

Reassembling of functional low immunogenic ICCs after single‐cell engineering. Genetically engineered cells were cultured for 7 d in a stirred bioreactor flask in the presence of collagen type VI to reassemble the ICCs. A, Representative microscopy images at different time points of the reassembling of the genetically engineered islets. B, Stirred bioreactor flasks containing genetically engineered ICCs‐derived cells during the reassembly of the ICCs. C, Representative microscopy image of the genetically engineered ICCs expressing GFP at the final stage of the islet reassembling. D, Illustrative microscopy image of native non‐engineered islets. Scale bar 100 µm. E, Reassembled ICCs were handpicked and incubated without (Negative Ctrl) or with 20 mmol/L glucose for 1 h at 37°C. Non‐engineered reassembled ICCs (Ctrl) or shRNA‐expressing reassembled ICCs were collected after glucose treatment, and total RNA was isolated for insulin transcript levels analysis. Graph shows insulin transcript levels of reassembled ICCs before and after stimulation with glucose. Insulin transcript levels were normalized to the endogenous control gene GAPDH. Graph shows means and standard deviations of three independent assays from all different ICCs. Statistical significances were calculated using one‐way ANOVA followed by Tukey's post hoc test. ***P < .001