Expansion and conversion of human pancreatic ductal cells into insulin-secreting endocrine cells.
Expansion and conversion of human pancreatic ductal cells into insulin-secreting endocrine cells.
复制标题
作者：
Lee J;Sugiyama T;Liu Y;Wang J;Gu X;Lei J;Markmann JF;Miyazaki S;Miyazaki J;Szot GL;Bottino R;Kim SK
胰岛β细胞功能不全是糖尿病发病机制的基础因此，利用可再生资源替代功能性β细胞是全世界努力的焦点。然而，在体外生产的后代分泌胰岛素响应生理线索从原代人细胞已被证明是难以捉摸的。在这里，我们描述了原代成人胰腺导管细胞的分离、扩增和转化为类似天然β细胞的后代。facs分类的成人导管细胞在培养中克隆扩增成球，同时保留导管特征。主要胰岛发育调节因子Neurog3、MafA、Pdx1和Pax6的表达将外分泌管细胞转化为具有标志性β细胞特性的内分泌后代，包括合成、加工和储存胰岛素的能力，以及在葡萄糖或其他去极化刺激下分泌胰岛素的能力。这些研究提供的证据表明，具有确定因子的可扩展人类胰腺细胞的遗传重编程可能作为糖尿病胰岛替代的一般策略。DOI: http://dx.doi.org/10.7554/eLife.00940.001糖尿病是一种可导致危险的高血糖水平的疾病，引起许多并发症，如心脏病、青光眼、皮肤病、肾病和神经损伤。在健康的个体中，胰腺中的β细胞会产生一种叫做胰岛素的激素，这种激素会刺激肝脏、肌肉和脂肪中的细胞从血液中吸收葡萄糖。然而，这个过程在糖尿病患者中被破坏，他们要么胰腺细胞太少（1型糖尿病），要么对胰岛素没有适当的反应（2型糖尿病）。所有1型糖尿病患者和一些2型糖尿病患者都必须定期注射胰岛素，但这并不总是能完全控制疾病。一些1型患者已经成功地接受了来自已故供体的β细胞移植治疗，但没有足够的供体器官使其成为常规。因此，全世界都在集中精力在实验室中从人类干细胞中生成胰岛素生成细胞。然而，以这种方式产生的细胞会产生肿瘤。现在，Lee等人已经证明，约占人体胰腺30%的导管细胞可以转化为能够产生和分泌胰岛素的细胞。从供体胰腺中获得的导管细胞首先从剩余组织中分离出来并在细胞培养中生长。然后用病毒来引入基因，重新编程导管细胞，使它们获得制造、加工和储存胰岛素的能力，并根据功能β细胞的葡萄糖特征释放胰岛素。除了为糖尿病的移植或细胞转化疗法提供潜在的细胞来源外，培养和维持人类胰腺导管细胞的能力可能使研究其他影响胰腺的疾病（包括胰腺炎、囊性纤维化和腺癌）变得更容易。DOI: http://dx.doi.org/10.7554/eLife.00940.002
Pancreatic islet β-cell insufficiency underlies pathogenesis of diabetes mellitus; thus, functional β-cell replacement from renewable sources is the focus of intensive worldwide effort. However, in vitro production of progeny that secrete insulin in response to physiological cues from primary human cells has proven elusive. Here we describe fractionation, expansion and conversion of primary adult human pancreatic ductal cells into progeny resembling native β-cells. FACS-sorted adult human ductal cells clonally expanded as spheres in culture, while retaining ductal characteristics. Expression of the cardinal islet developmental regulators Neurog3, MafA, Pdx1 and Pax6 converted exocrine duct cells into endocrine progeny with hallmark β-cell properties, including the ability to synthesize, process and store insulin, and secrete it in response to glucose or other depolarizing stimuli. These studies provide evidence that genetic reprogramming of expandable human pancreatic cells with defined factors may serve as a general strategy for islet replacement in diabetes. DOI: http://dx.doi.org/10.7554/eLife.00940.001 Diabetes mellitus is a disease that can lead to dangerously high blood sugar levels, causing numerous complications such as heart disease, glaucoma, skin disorders, kidney disease, and nerve damage. In healthy individuals, beta cells in the pancreas produce a hormone called insulin, which stimulates cells in the liver, muscles and fat to take up glucose from the blood. However, this process is disrupted in people with diabetes, who either have too few pancreatic beta cells (type 1 diabetes) or do not respond appropriately to insulin (type 2 diabetes). All patients with type 1 diabetes, and some with type 2, must inject themselves regularly with insulin, but this does not always fully control the disease. Some type 1 patients have been successfully treated with beta cells transplanted from deceased donors, but there are not enough donor organs available for this to become routine. Thus, intensive efforts worldwide are focused on generating insulin-producing cells in the lab from human stem cells. However, the cells produced in this way can give rise to tumors. Now, Lee et al. have shown that duct cells, which make up about 30% of the human pancreas, can be converted into cells capable of producing and secreting insulin. Ductal cells obtained from donor pancreases were first separated from the remaining tissue and grown in cell culture. Viruses were then used to introduce genes that reprogrammed the ductal cells so that they acquired the ability to make, process and store insulin, and to release it in response to glucose—hallmark features of functional beta cells. As well as providing a potential source of cells for use in transplant or cell conversion therapies for diabetes, the ability to grow and maintain human pancreatic ductal cells in culture may make it easier to study other diseases that affect the pancreas, including pancreatitis, cystic fibrosis, and adenocarcinoma. DOI: http://dx.doi.org/10.7554/eLife.00940.002