Investigation of medium perfusion through scaffold-free tissue constructs using endothelial cell-covered spheroids in vitro
Investigation of medium perfusion through scaffold-free tissue constructs using endothelial cell-covered spheroids in vitro
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DOI：
10.1016/j.bej.2010.04.004
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发表时间：
2010-07-15
影响因子：
3.9
通讯作者：
Kajiwara, Toshihisa
中科院分区：
文献类型：
--
作者：
Inamori, Masakazu;Mizumoto, Hiroshi;Kajiwara, Toshihisa
通过组装内皮细胞覆盖的球体形成无支架组织结构，并利用流体静压驱动培养电路研究通过该组织结构的介质灌流。将包裹有人脐静脉内皮细胞(HUVECs)的原代大鼠肝细胞球体组装在直径为2 mm的圆柱形培养腔中，然后通过组装的球体灌流培养48h，测量在灌流培养过程中流过培养腔的介质流量，在最初几小时内有所下降，然后根据培养腔中球体的数量增加或保持在较低的水平。组织化学分析显示，当从2×10(5)核球体培养时，单个组织结构由球体融合形成，球体之间的边界消失。此外，在组织结构的中心在几个位置发现了一个活细胞区域。4×10(5)核培养的球体之间粘附性差。培养的组织结构中的总核密度估计约为HUVEC覆盖的肝细胞球体的一半。本研究证明了通过组装内皮细胞覆盖的球体通过无支架组织结构进行介质灌流的可能性，有望在体外进行大型组织结构的培养。(C)2010年爱思唯尔BM。版权所有。
A scaffold-free tissue construct was formed by assembling endothelial cell-covered spheroids, and medium perfusion through the tissue construct was investigated using hydrostatic pressure-driven culture circuit. Primary rat hepatocyte spheroids covered by human umbilical vein endothelial cells (HUVECs) were assembled in culture chambers with a cylindrical culture space of 2 mm in diameter, and then medium was perfused through the assembled spheroids for 48 h. The medium flow rate through the culture chamber was measured over the perfusion culture time, which decreased during the first several hours, then increased or remained low depending on the amount of spheroids in the culture chamber. Histochemical analyses showed single tissue construct formation by spheroid fusion when cultured from 2 x 10(5) nuclei spheroids, with the loss of boundaries between the spheroids. Moreover, a viable cell region was found at the center of the tissue construct in several locations. Poor adhesion was found between spheroids cultured from 4 x 10(5) nuclei spheroids. The total nuclei density in cultured tissue constructs was estimated to be about half of that in HUVEC-covered hepatocyte spheroids.This study demonstrated the possibility of medium perfusion through scaffold-free tissue constructs by assembling endothelial cell-covered spheroids, promising for a large tissue construct culture in vitro. (C) 2010 Elsevier BM. All rights reserved.