Microenvironment-regulated gene expression, morphology, and in vivo performance of mouse pancreatic β-cells
2013
Chen, Pei-Yu | Wu, Chang-Chin | Lu, Dai-Hua | Sumi, Shoichiro | Lin, Feng-Huei | Yang, Kai-Chiang
Cell behavior is determined by intrinsic characteristics and complex interactions with microenvironments. This study demonstrated the performance of a murine pancreatic β-cell line, MIN-6, cultured on tissue-culture polystyrene (TCPS), gelatin, type I collagen, and type IV collagen dishes. MIN-6 cells aggregated as clusters on gelatin, type I collagen, and type IV collagen, which was different from the epithelial morphology of cells grown on TCPS. The diameter and survival rate of aggregated cells did not differ significantly regardless of whether the cells were grown on gelatin or type I collagen, while smaller clusters were observed on type IV collagen. Compared with the monolayers on TCPS, the clusters had a higher insulin stimulation index. The mRNA expression levels of Ins1, Pdx-1, NeuroD1 and connexin 36 were upregulated in clusters relative to monolayers. Conversely, E-cadherin and MafA were downregulated when cells were grown on type IV collagen. Monolayers or cell aggregates grown on type IV collagen were subsequently transplanted into diabetic C57BL/6 mice. Animals that received both monolayers and clusters had decreased blood glucose levels and regained body weight. However, the area under curve for the intraperitoneal glucose tolerance test showed that clusters exhibited superior in vivo performance. This study reveals that a type IV collagen substrate promotes β-cell clustering, regulates gene expression and enhances in vivo performance.
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