Isolation of major pancreatic cell types and long-term culture-initiating cells using novel human surface markers
Isolation of major pancreatic cell types and long-term culture-initiating cells using novel human surface markers
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DOI：
10.1016/j.scr.2008.04.001
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发表时间：
2008-09-01
影响因子：
1.2
通讯作者：
Grompe, Markus
中科院分区：
文献类型：
--
作者：
Dorrell, Craig;Abraham, Stephanie L.;Grompe, Markus
我们开发了一组新的细胞表面标记物，用于分离和研究人胰腺的所有主要细胞类型。用完整的或离散的人胰岛对Balb/C小鼠进行消减免疫后筛选杂交瘤，并通过免疫组织化学和流式细胞术评估细胞类型特异性和细胞表面反应性。通过胰岛（泛内分泌或α特异性）和非胰岛胰腺细胞亚群（外分泌和导管）表面抗原的特异性结合来鉴定抗体。这些抗体单独或组合使用，通过流式细胞术从原代人胰腺中分离出α、β、外分泌或导管细胞群，并表征人胰岛制剂的详细细胞组成。它们还被用于表明人胰岛扩增培养物起源于非内分泌细胞，并且通过亚群分选以及转录因子Pdx - 1和ngn3的过表达，胰岛素表达水平可提高至正常胰岛细胞的1%，这比该培养系统先前的结果有所改进。这些方法允许对具有细胞治疗潜力的功能不同的胰腺细胞群进行分析和分离。（c）2008由爱思唯尔B.V.出版
We have developed a novel panel of cell-surface markers for the isolation and study of all major cell types of the human pancreas. Hybridomas were selected after subtractive immunization of Balb/C mice with intact or dissociated human islets and assessed for cell-type specificity and cell-surface reactivity by immunohistochemistry and flow cytometry. Antibodies were identified by specific binding of surface antigens on islet (panendocrine or alpha-specific) and nonislet pancreatic cell subsets (exocrine and duct). These antibodies were used individually or in combination to isolate populations of alpha, beta, exocrine, or duct cells from primary human pancreas by FACS and to characterize the detailed cell composition of human islet preparations. They were also employed to show that human islet expansion cultures originated from nonendocrine cells and that insulin expression levels could be increased to up to 1% of normal islet cells by subpopulation sorting and overexpression of the transcription factors Pdx-1 and ngn3, an improvement over previous results with this culture system. These methods permit the analysis and isolation of functionally distinct pancreatic cell populations with potential for cell therapy. (c) 2008 Published by Elsevier B.V.