Formation of microvascular networks in vitro
Formation of microvascular networks in vitro
复制标题
DOI：
10.1038/nprot.2013.110
复制
发表时间：
2013-09-01
期刊：
影响因子：
14.8
通讯作者：
Stroock, Abraham D.
中科院分区：
文献类型：
--
作者：
Morgan, John P.;Delnero, Peter F.;Stroock, Abraham D.
该协议描述了如何形成具有显式内皮化微血管的 3D 细胞培养物。该方法在生物可重塑水凝胶（I 型胶原）中形成完全封闭的可灌注血管。该协议使用微加工来实现用户定义的血管网络和微流体灌注的几何形状，以控制质量传递和血流动力学。这些微血管网络 (mVN) 允许内皮细胞进行数周培养，或与管腔外空间的实质或组织细胞共培养。该平台能够对活体工程组织进行实时荧光成像、固定培养物的原位共焦荧光以及组织切片的透射电子显微镜 (TEM) 成像。该协议能够研究基础血管和血液生物学，为肿瘤血管生成或血栓形成等疾病提供模型，并作为构建再生医学预血管化组织的起点。经过一次性微加工步骤后，系统可以在不到 1 天内组装完毕，实验可以运行数周。
This protocol describes how to form a 3D cell culture with explicit, endothelialized microvessels. The approach leads to fully enclosed, perfusable vessels in a bioremodelable hydrogel (type I collagen). The protocol uses microfabrication to enable user-defined geometries of the vascular network and microfluidic perfusion to control mass transfer and hemodynamic forces. These microvascular networks (mVNs) allow for multiweek cultures of endothelial cells or cocultures with parenchymal or tissue cells in the extra-lumen space. The platform enables real-time fluorescence imaging of living engineered tissues, in situ confocal fluorescence of fixed cultures and transmission electron microscopy (TEM) imaging of histological sections. This protocol enables studies of basic vascular and blood biology, provides a model for diseases such as tumor angiogenesis or thrombosis and serves as a starting point for constructing prevascularized tissues for regenerative medicine. After one-time microfabrication steps, the system can be assembled in less than 1 d and experiments can run for weeks.