In vitro generation of insulin-producing beta cells from adult exocrine pancreatic cells
In vitro generation of insulin-producing beta cells from adult exocrine pancreatic cells
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DOI：
10.1007/s00125-004-1606-1
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发表时间：
2005-01-01
期刊：
影响因子：
8.2
通讯作者：
Bouwens, L
中科院分区：
文献类型：
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作者：
Baeyens, L;De Breuck, S;Bouwens, L
目的/假设：移植来自捐赠者的产生胰岛素的β细胞可以治愈糖尿病，但数量不足。在这项研究中，我们研究了在生长因子存在下培养的成年大鼠外分泌细胞产生胰岛素产生细胞的可能性。方法：分离大鼠外分泌胰腺细胞，用表皮生长因子(EGF)和白血病抑制因子(LIF)进行体外培养。采用免疫细胞化学、DNA检测和放射免疫分析等方法进行分析。将细胞移植到经四氧嘧啶(70 mg/kg)处理的裸鼠体内，连续21天监测血糖。第15天行肾切除。结果：在3d培养期间，加入LIF+EGF的培养液中，β细胞质量增加了11倍。这不能归因于污染的β细胞的有丝分裂活性非常低。此外，当污染的β细胞最初被四氧嘧啶破坏时，这种影响更加明显。新形成的细胞在葡萄糖的作用下分泌胰岛素，并对C-肽-I、PDX-1和GLUT-2呈免疫反应，这些是成熟β细胞的特征。电子显微镜显示，它们还含有胰岛素免疫反应分泌颗粒。部分胰岛素阳性细胞呈淀粉酶和细胞角蛋白-20免疫反应阳性，或双核，为外分泌细胞的特征。当将这些细胞移植到四氧嘧啶糖尿病小鼠身上时，这些细胞能够恢复正常的血糖，在移植物移除后，高血糖复发。结论/解释：我们的研究表明，外分泌组织通过转分化可以产生功能性的β细胞，因此可能为β细胞治疗提供一个新的视角。
Aims/hypothesis: Transplantation of insulinproducing beta cells from donors can cure diabetes, but they are available in insufficient quantities. In this study, we investigated the possibility of generating insulinproducing cells from adult rat exocrine cells cultured in the presence of growth factors. Methods: Rat exocrine pancreatic cells were isolated and treated in vitro with epidermal growth factor ( EGF) and leukaemia inhibitory factor ( LIF). Analysis was performed by immunocytochemistry, DNA measurement and radioimmunoassay. Cells were transplanted to alloxan- treated ( 70 mg/ kg) nude mice and glycaemia was monitored for 21 days. Nephrectomy was performed on day 15. Results: In a 3- day culture period, addition of LIF plus EGF to the medium resulted in an 11- fold increase of the beta cell mass. This could not be attributed to the very low mitotic activity of contaminating beta cells. Furthermore, when contaminating beta cells were initially destroyed with alloxan, this effect was even more pronounced. The newly formed cells secreted insulin in response to glucose and were immunoreactive for C- peptide- I, Pdx- 1 and GLUT-2, which are characteristics of mature beta cells. Electron microscopy showed that they also contained insulin-immunoreactive secretory granules. Some insulin-positive cells were immunoreactive for amylase and cytokeratin-20, or were binucleated, which are characteristics of exocrine cells. The cells were able to restore normoglycaemia when transplanted to alloxan- diabetic mice, and hyperglycaemia recurred upon removal of the graft. Conclusions/interpretation: Our study shows that functional beta cells can be generated from exocrine tissue by transdifferentiation and thereby may offer a new perspective for beta cell therapy.