Mfopou Josué K, Houbracken Isabelle, Wauters Elke, Mathijs Iris, Song Imane, Himpe Eddy, Baldan Jonathan, Heimberg Harry, Bouwens Luc
Cell Differentiation Unit, Diabetes Research Center, Vrije Universiteit Brussel (VUB), Brussels, BE 1090, Belgium.
Beta Cell Neogenesis, Diabetes Research Center, Vrije Universiteit Brussel (VUB), Brussels, BE 1090, Belgium.
Biosci Rep. 2016 May 6;36(3). doi: 10.1042/BSR20150259. Print 2016 Jun.
The regenerative medicine field is expanding with great successes in laboratory and preclinical settings. Pancreatic acinar cells in diabetic mice were recently converted into β-cells by treatment with ciliary neurotrophic factor (CNTF) and epidermal growth factor (EGF). This suggests that human acinar cells might become a cornerstone for diabetes cell therapy in the future, if they can also be converted into glucose-responsive insulin-producing cells. Presently, studying pancreatic acinar cell biology in vitro is limited by their high plasticity, as they rapidly lose their phenotype and spontaneously transdifferentiate to a duct-like phenotype in culture. We questioned whether human pancreatic acinar cell phenotype could be preserved in vitro by physico-chemical manipulations and whether this could be valuable in the study of β-cell neogenesis. We found that culture at low temperature (4°C) resulted in the maintenance of morphological and molecular acinar cell characteristics. Specifically, chilled acinar cells did not form the spherical clusters observed in controls (culture at 37°C), and they maintained high levels of acinar-specific transcripts and proteins. Five-day chilled acinar cells still transdifferentiated into duct-like cells upon transfer to 37°C. Moreover, adenoviral-mediated gene transfer evidenced an active Amylase promoter in the 7-day chilled acinar cells, and transduction performed in chilled conditions improved acinar cell labelling. Together, our findings indicate the maintenance of human pancreatic acinar cell phenotype at low temperature and the possibility to efficiently label acinar cells, which opens new perspectives for the study of human acinar-to-β-cell transdifferentiation.
再生医学领域正在不断扩展,在实验室和临床前研究中都取得了巨大成功。最近,通过睫状神经营养因子(CNTF)和表皮生长因子(EGF)处理,糖尿病小鼠的胰腺腺泡细胞被转化为β细胞。这表明,如果人类腺泡细胞也能被转化为对葡萄糖有反应的胰岛素产生细胞,那么它们可能会成为未来糖尿病细胞治疗的基石。目前,体外研究胰腺腺泡细胞生物学受到其高可塑性的限制,因为它们在培养中会迅速失去其表型并自发转分化为导管样表型。我们质疑人类胰腺腺泡细胞表型是否可以通过物理化学操作在体外得以保留,以及这在β细胞新生研究中是否有价值。我们发现,在低温(4°C)下培养可维持腺泡细胞的形态和分子特征。具体而言,冷冻的腺泡细胞不会形成在对照组(37°C培养)中观察到的球形簇,并且它们维持高水平的腺泡特异性转录本和蛋白质。五天的冷冻腺泡细胞在转移到37°C后仍会转分化为导管样细胞。此外,腺病毒介导的基因转移证明在7天的冷冻腺泡细胞中有活跃的淀粉酶启动子,并且在冷冻条件下进行的转导改善了腺泡细胞标记。总之,我们的研究结果表明人类胰腺腺泡细胞表型在低温下得以维持,并且有可能有效地标记腺泡细胞,这为研究人类腺泡细胞向β细胞的转分化开辟了新的前景。