Isolation and characterization of centroacinar/terminal ductal progenitor cells in adult mouse pancreas
Isolation and characterization of centroacinar/terminal ductal progenitor cells in adult mouse pancreas
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DOI：
10.1073/pnas.0912589107
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发表时间：
2010-01-05
影响因子：
11.1
通讯作者：
Leach, Steven D.
中科院分区：
文献类型：
--
作者：
Rovira, Meritxell;Scott, Sherri-Gae;Leach, Steven D.
成年脊椎动物胰腺中是否存在专门的祖细胞的问题仍然存在争议。中央腺泡细胞和终末导管（CA/TD）细胞位于外周腺泡细胞和相邻导管上皮之间的交界处，并且经常被包括在被提出作为候选胰腺祖细胞的细胞类型中。然而，这些细胞以前没有以允许正式评估其祖细胞能力的方式分离。我们已经发现，一个子集的成人CA/TD细胞的特点是高水平的ALDH 1酶活性，与高水平表达的Aldh 1a 1和Aldh 1a 7。这允许使用荧光ALDH 1底物通过FACS分离它们。FACS分离的CA/TD细胞相对耗尽与分化的胰腺细胞类型相关的转录物。相反，它们明显富集编码Sca 1、Sdf 1、c-Met、巢蛋白和Sox 9的转录物，这些标记物先前与胚胎胰腺和其他组织中的祖细胞群体相关。FACS分选的CA/TD细胞独特地能够在悬浮培养物中形成自我更新的“胰腺球”，甚至当以克隆密度铺板时。这些球体显示出自发内分泌和外分泌分化以及葡萄糖响应性胰岛素分泌的能力。此外，当注入培养的胚胎背胰芽时，这些成体细胞显示出独特的能力，有助于胚胎内分泌和外分泌谱系。最后，这些细胞在慢性上皮损伤的情况下表现出显著的扩增。这些发现表明，CA/TD细胞确实能够祖细胞功能，并可能有助于维持组织稳态在成年小鼠胰腺。
The question of whether dedicated progenitor cells exist in adult vertebrate pancreas remains controversial. Centroacinar cells and terminal duct (CA/TD) cells lie at the junction between peripheral acinar cells and the adjacent ductal epithelium, and are frequently included among cell types proposed as candidate pancreatic progenitors. However these cells have not previously been isolated in a manner that allows formal assessment of their progenitor capacities. We have found that a subset of adult CA/TD cells are characterized by high levels of ALDH1 enzymatic activity, related to high-level expression of both Aldh1a1 and Aldh1a7. This allows their isolation by FACS using a fluorogenic ALDH1 substrate. FACS-isolated CA/TD cells are relatively depleted of transcripts associated with differentiated pancreatic cell types. In contrast, they are markedly enriched for transcripts encoding Sca1, Sdf1, c-Met, Nestin, and Sox9, markers previously associated with progenitor populations in embryonic pancreas and other tissues. FACS-sorted CA/TD cells are uniquely able to form self-renewing "pancreatospheres" in suspension culture, even when plated at clonal density. These spheres display a capacity for spontaneous endocrine and exocrine differentiation, as well as glucose-responsive insulin secretion. In addition, when injected into cultured embryonic dorsal pancreatic buds, these adult cells display a unique capacity to contribute to both the embryonic endocrine and exocrine lineages. Finally, these cells demonstrate dramatic expansion in the setting of chronic epithelial injury. These findings suggest that CA/TD cells are indeed capable of progenitor function and may contribute to the maintenance of tissue homeostasis in adult mouse pancreas.