Characterization of pancreatic stem cells derived from adult human pancreas ducts by fluorescence activated cell sorting

AIM： To isolate putative pancreatic stem cells （PSCs） from human adult tissues of pancreas duct using serumfree, conditioned medium. The characterization of surface phenotype of these PSCs was analyzed by flow cytometry. The potential for pancreatic lineage and the capability of β-cell differentiation in these PSCs were evaluated as well. METHODS： By using serum-free medium supplemented with essential growth factors, we attempted to isolate the putative PSCs which has been reported to express nestin and pdx-1. The MatrigelTM was employed to evaluate the differential capacity of isolated cells. Dithizone staining, insulin content/secretion measurement, and immunohistochemistry staining were used to monitor the differentiation. Fluorescence activated cell sorting （FACS） was used to detect the phenotypic markers of putative PSCs. RESULTS： A monolayer of spindle-like cells was cultivated. The putative PSCs expressed pdx-1 and nestin. They were also able to differentiate into insulin-, glucagon-, and somatostatin-positive cells. The spectrum of phenotypic markers in PSCs was investigated; a similarity was revealed when using human bone marrow-derived stem cells as the comparative experiment, such as CD29, CD44, CD49, CD50, CD51, CD62E, PDGFR-α, CD73 （SH2）, CD81, CD105（SH3）. CONCLUSION： In this study, we successfully isolated PSCs from adult human pancreatic duct by using serumfree medium. These PSCs not only expressed nestin and pdx-1 but also exhibited markers attributable to mesenchymal stem cells. Although work is needed to elucidate the role of these cells, the application of these PSCs might be therapeutic strategies for diabetes mellitus.

瞄准：用没有浆液的、调节媒介从胰管的人的成年纸巾孤立通常认为的胰腺的干细胞(PSC ) 。这些 PSC 的表面显型的描述被流动血细胞计数分析。为胰腺的系和在这些 PSC 的贝它房间区别的能力的潜力也被评估。方法：由使用与必要生长因素补充的没有浆液的媒介，我们试图孤立被报导了表示巢在的通常认为的 PSC 和 pdx-1。Matrigel (TM ) 被采用评估孤立的房间的微分能力。染色的染色的双硫腙，胰岛素内容 / 分泌物测量，和免疫组织化学被用来监视区别。荧光(FACS ) 激活的房间排序被用来检测通常认为的 PSC 的表型的标记。结果：像锭子的房间的单层被栽培。通常认为的 PSC 表示了 pdx-1 和巢在里面。他们也能区分进胰岛素 -- ， glucagon- ，和 somatostatin 积极的房间。在 PSC 的表型的标记的光谱被调查；当用人的骨头导出髓的干细胞作为比较级试验时，类似被揭示，例如 CD29， CD44， CD49，半数治愈量， CD51， CD62E， PDGFR-alpha， CD73 (SH2 ) ， CD81， CD105 (SH3 ) 。结论：在这研究，我们成功地由使用没有浆液的媒介从成年的人的胰腺的管孤立 PSC。这些 PSC 不仅表示的巢在和 pdx-1 而且对间充质的干细胞可归因的展出标记。尽管工作被需要阐明这些房间的角色，这些 PSC 的申请可能是为糖尿病的治疗学的策略。