Endoglin (CD105) is not a specific selection marker for endothelial cells in human islets of Langerhans
Endoglin (CD105) is not a specific selection marker for endothelial cells in human islets of Langerhans
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DOI：
10.1007/s00125-012-2763-2
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发表时间：
2012
期刊：
影响因子：
8.2
通讯作者：
C. Wheeler-Jones;C. Clarkin;C. Farrar;Pam Dhadda;P. Chagastelles;N. Nardi;P. Jones
中科院分区：
文献类型：
--
作者：
C. Wheeler-Jones;C. Clarkin;C. Farrar;Pam Dhadda;P. Chagastelles;N. Nardi;P. Jones
致编辑：我们饶有兴趣地阅读了 Favaro 等人最近的报告 [1]，该报告表明，从人胰岛中分离出的微血管内皮细胞 (MEC) 可以通过胃肠道肽来保护，免受高血糖诱导的细胞凋亡。在这项研究中，与之前的几项研究一样 [2-5]，MEC 在体外从使用抗 CD105 免疫珠从分散的人胰岛细胞中分离出来的群体中扩增。 CD105（也称为内皮糖蛋白）是一种膜糖蛋白，与转化生长因子 (TGF)-β 受体共同作用，作为 TGF-β 家族成员（包括激活素、TGF-β1/β3 和骨形态发生蛋白）的辅助结合蛋白。大血管和微血管内皮细胞 (EC) 肯定是 CD105 阳性，但这种辅助受体也在一系列其他细胞类型中高度表达，包括骨髓和组织源性间充质基质细胞 (MSC) 样群。据此，我们最近从人类胰岛中分离并在体外扩增了一群表达 CD105 的细胞，这些细胞显然不是 MEC，但它们与基于 CD105 表达进行免疫分离的其他细胞具有相同的表型特征，并被鉴定为用于体外研究的胰岛 MEC [1-5]。我们的细胞群基本上按照 MSC 的描述进行分离和扩增 [6]。简而言之，通过在 0.02%（重量/体积）EDTA 溶液中孵育并研磨来分离人类胰岛（从伦敦国王学院人类胰岛分离装置获得，并获得适当的伦理批准）。将细胞重悬于补充有 L-谷氨酰胺 (2 mM)、青霉素 (100 U/ml)、链霉素 (172 μmol/l) 和 FCS (10% vol./vol.) 的 Dulbecco’s Modified Eagle’s 培养基中，接种在六孔组织培养皿中，并在含有 5%/95% CO2/空气的潮湿气氛中于 37°C 下孵育。 24小时后更换培养基，去除非贴壁细胞，当贴壁培养物达到汇合时，将它们进行胰蛋白酶消化并传代培养到组织培养瓶中。这些细胞可能是贴壁胰岛基质细胞的异质群体，包括成纤维细胞、肌成纤维细胞和周细胞，以及具有分化潜力的间充质干细胞。据此，这些细胞具有许多与 MSC 相关的属性 [6, 7]，而不是 MEC，如图 1 所示。它们没有表现出单层 EC 的典型鹅卵石形态（图 1a-c），但具有成纤维细胞样纺锤体
To the Editor: We read with interest the recent report by Favaro et al [1] suggesting that microvascular endothelial cells (MECs) isolated from human islets of Langerhans could be protected from hyperglycaemia-induced apoptosis by gastrointestinal peptides. In this study, as in several previous studies [2–5], the MECs were expanded in vitro from a population that had been isolated from dispersed human islet cells using anti-CD105 immunobeads. CD105 (also known as endoglin) is a membrane glycoprotein that acts with transforming growth factor (TGF)-β receptors as an auxiliary binding protein for members of the TGF-β family, including activin, TGF-β1/β3 and bone morphogenic proteins. Macro-and microvascular endothelial cells (ECs) are certainly CD105-positive, but this accessory receptor is also highly expressed by a range of other cell types including bone marrow-and tissue-derived mesenchymal stromal cell (MSC)-like populations. In accordance with this, we have recently isolated from human islets, and expanded in vitro, a population of CD105-expressing cells that are clearly not MECs, but which share phenotypic characteristics with other cells that have been immuno-isolated on the basis of CD105 expression and identified as islet MECs for in vitro studies [1–5].Our cell population was isolated and expanded essentially as described for MSCs [6]. Briefly, human islets (obtained from the King’s College London Human Islet Isolation Unit, with appropriate ethics approval) were dissociated by incubation in 0.02%(wt/vol.) EDTA solution with trituration. Cells were resuspended in Dulbecco’s Modified Eagle’s Medium supplemented with L-glutamine (2 mM), penicillin (100 U/ml), streptomycin (172 μmol/l) and FCS (10% vol./vol.), seeded in six-well tissue culture dishes and incubated at 37 C in a humidified atmosphere containing 5%/95% CO2/air. The medium was changed after 24 h, with removal of non-adherent cells, and when adherent cultures reached confluence, they were trypsinised and subcultured into tissue culture flasks. These cells are likely to be a heterogeneous population of adherent islet stromal cells, including fibroblasts, myofibroblasts and pericytes, as well as MSCs with differentiation potential. In accordance with this, the cells possessed many of the attributes associated with MSCs [6, 7], rather than MECs, as shown in Fig. 1. They did not exhibit the typical cobble-stone morphology of monolayer ECs (Fig. 1a–c), but had a fibroblast-like spindle