Ye F, Duvillié B, Scharfmann R
Faculty Necker, INSERM E363, 156 Rue de Vaugirard, 75015 Paris, France.
Diabetologia. 2005 Feb;48(2):277-81. doi: 10.1007/s00125-004-1638-6. Epub 2005 Feb 3.
AIMS/HYPOTHESIS: The fibroblast growth factor (FGF) family consists of 22 members. In rodents, several FGFs are expressed in the pancreas, where they participate in epithelial-mesenchymal interactions. Our objective was to describe the pattern of expression of FGFs in the human embryonic pancreas and to analyse their effect on pancreas development.
The expression of FGFs was analysed by RT-PCR. To investigate the cell types expressing FGF7 and FGF10, we separated epithelial from mesenchymal cells using immunomagnetic beads linked to E-cadherin antibodies and performed real-time PCR. The effect of FGF7 and FGF10 on proliferation of human embryonic pancreatic epithelial cells was evaluated in vitro by measuring BrdU incorporation.
We found that different FGFs are expressed in the human embryonic pancreas, and we focused on FGF7 and FGF10. We defined a new approach to separating epithelial cells (containing the pancreatic progenitor cells) from mesenchymal cells. This allowed us to demonstrate that human embryonic pancreatic mesenchymal cells express both FGF7 and FGF10. We next demonstrated that FGF7 and FGF10 were able to induce the proliferation of the epithelial cells in vitro.
CONCLUSION/INTERPRETATION: These findings indicate that it is now possible to efficiently separate human embryonic pancreatic epithelial from mesenchymal cells, an important step to characterize and expand progenitor cells. This method allowed us to demonstrate that human embryonic pancreatic mesenchyme expresses FGF7 and FGF10 that act on epithelial cells to activate their proliferation. Such growth factors could thus be used to expand human embryonic pancreatic epithelial cells.
目的/假设:成纤维细胞生长因子(FGF)家族由22个成员组成。在啮齿动物中,几种FGF在胰腺中表达,它们参与上皮-间充质相互作用。我们的目的是描述FGF在人类胚胎胰腺中的表达模式,并分析它们对胰腺发育的影响。
通过逆转录聚合酶链反应(RT-PCR)分析FGF的表达。为了研究表达FGF7和FGF10的细胞类型,我们使用与E-钙黏蛋白抗体相连的免疫磁珠从间充质细胞中分离出上皮细胞,并进行实时聚合酶链反应。通过测量5-溴脱氧尿嘧啶核苷(BrdU)掺入量,在体外评估FGF7和FGF10对人类胚胎胰腺上皮细胞增殖的影响。
我们发现不同的FGF在人类胚胎胰腺中表达,并且我们重点研究了FGF7和FGF10。我们定义了一种从间充质细胞中分离上皮细胞(包含胰腺祖细胞)的新方法。这使我们能够证明人类胚胎胰腺间充质细胞同时表达FGF7和FGF10。接下来我们证明FGF7和FGF10能够在体外诱导上皮细胞增殖。
结论/解读:这些发现表明现在有可能有效地从间充质细胞中分离出人类胚胎胰腺上皮细胞,这是鉴定和扩增祖细胞的重要一步。这种方法使我们能够证明人类胚胎胰腺间充质表达FGF7和FGF10,它们作用于上皮细胞以激活其增殖。因此,此类生长因子可用于扩增人类胚胎胰腺上皮细胞。