INSERM U845, Research Center Growth and Signalling, Faculté de Médecine Cochin, Université Paris Descartes, Paris, France.
Diabetes. 2013 Oct;62(10):3479-87. doi: 10.2337/db13-0554. Epub 2013 Jul 8.
While sporadic human genetic studies have permitted some comparisons between rodent and human pancreatic development, the lack of a robust experimental system has not permitted detailed examination of human pancreatic development. We previously developed a xenograft model of immature human fetal pancreas grafted under the kidney capsule of immune-incompetent mice, which allowed the development of human pancreatic β-cells. Here, we compared the development of human and murine fetal pancreatic grafts either under skeletal muscle epimysium or under the renal capsule. We demonstrated that human pancreatic β-cell development occurs more slowly (weeks) than murine pancreas (days) both by differentiation of pancreatic progenitors and by proliferation of developing β-cells. The superficial location of the skeletal muscle graft and its easier access permitted in vivo lentivirus-mediated gene transfer with a green fluorescent protein-labeled construct under control of the insulin or elastase gene promoter, which targeted β-cells and nonendocrine cells, respectively. This model of engraftment under the skeletal muscle epimysium is a new approach for longitudinal studies, which allows localized manipulation to determine the regulation of human pancreatic development.
虽然零星的人类遗传学研究允许在啮齿动物和人类胰腺发育之间进行一些比较,但缺乏强大的实验系统使得无法详细检查人类胰腺发育。我们之前开发了一种不成熟的人类胎儿胰腺异种移植模型,将其移植到免疫功能低下的小鼠肾脏包膜下,从而允许人类胰腺β细胞的发育。在这里,我们比较了在骨骼肌外膜下或肾脏包膜下的人胎和鼠胎胰腺移植物的发育情况。我们证明,人类胰腺β细胞的发育速度比鼠胰腺(数天)慢(数周),这是通过胰腺祖细胞的分化和发育中的β细胞的增殖来实现的。骨骼肌移植物的浅表位置及其更容易接近,允许使用绿色荧光蛋白标记的构建体进行体内慢病毒介导的基因转移,该构建体受胰岛素或弹性蛋白酶基因启动子的控制,分别靶向β细胞和非内分泌细胞。这种在骨骼肌外膜下的植入模型是一种新的纵向研究方法,允许局部操作来确定人类胰腺发育的调控。