Upcyte® Microvascular Endothelial Cells Repopulate Decellularized Scaffold
Upcyte® Microvascular Endothelial Cells Repopulate Decellularized Scaffold
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DOI：
10.1089/ten.tec.2011.0723
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发表时间：
2013-01-01
影响因子：
3
通讯作者：
Walles, Heike
中科院分区：
文献类型：
--
作者：
Scheller, Katharina;Dally, Iris;Walles, Heike
组织工程学中的一个普遍问题是血液供应不足，以保证组织细胞的存活和生理组织功能。为了解决这一局限性，我们开发了一种体外血管形成模型，在该模型中，脱细胞的猪小肠段代表胶原基质(生物血管支架[BioVaSc])中的毛细血管网络，再种植微血管内皮细胞(MvECs)。然而，由于微血管内皮细胞的供应总体上是有限的，并且随着这些细胞的快速去分化，我们应用了一种新的技术，该技术允许产生大量具有增殖能力的准原代细胞，同时保持其分化的功能。与原代细胞相比，这些所谓的上细胞微血管内皮细胞生长了额外的15个群体倍增(PD)。上皮微血管内皮细胞保持内皮特性，如von Willebrandt因子(VWF)、CD31和内皮型一氧化氮合酶(ENOS)的表达，以及Ulex uropaeus凝集素I染色阳性。上层细胞微血管内皮细胞也保留了生物学功能，如管状形成、细胞迁移和低密度脂蛋白(LDL)摄取，这些在PD27后仍然明显。用四甲基偶氮唑盐和活/死染色的初步实验表明，上层细胞的微血管内皮细胞重新填充了BioVaSc支架。与常规培养一样，这些细胞在定制的生物反应器系统中也表达关键的内皮分子(vWF、CD31和eNOS)，即使在延长14天后也是如此。上皮微血管内皮细胞和BioVaSc的结合代表了一种新的、有前途的血管生物反应器模型，该模型可以更好地反映器官，如肝脏。
A general problem in tissue engineering is the poor and insufficient blood supply to guarantee tissue cell survival as well as physiological tissue function. To address this limitation, we have developed an in vitro vascularization model in which a decellularized porcine small bowl segment, representing a capillary network within a collagen matrix (biological vascularized scaffold [BioVaSc]), is reseeded with microvascular endothelial cells (mvECs). However, since the supply of mvECs is limited, in general, and as these cells rapidly dedifferentiate, we have applied a novel technology, which allows the generation of large batches of quasi-primary cells with the ability to proliferate, whilst maintaining their differentiated functionality. These so called upcyte mvECs grew for an additional 15 population doublings (PDs) compared to primary cells. Upcyte mvECs retained endothelial characteristics, such as von Willebrandt Factor (vWF), CD31 and endothelial nitric oxide synthase (eNOS) expression, as well as positive Ulex europaeus agglutinin I staining. Upcyte mvECs also retained biological functionality such as tube formation, cell migration, and low density lipoprotein (LDL) uptake, which were still evident after PD27. Initial experiments using MTT and Live/Dead staining indicate that upcyte mvECs repopulate the BioVaSc Scaffold. As with conventional cultures, these cells also express key endothelial molecules (vWF, CD31, and eNOS) in a custom-made bioreactor system even after a prolonged period of 14 days. The combination of upcyte mvECs and the BioVaSc represents a novel and promising approach toward vascularizing bioreactor models which can better reflect organs, such as the liver.