Development of a scaffoldless three-dimensional engineered nerve using a nerve-fibroblast co-culture.
Development of a scaffoldless three-dimensional engineered nerve using a nerve-fibroblast co-culture.
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DOI：
10.1007/s11626-009-9260-z
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发表时间：
2010-05
影响因子：
2.1
通讯作者：
Larkin, Lisa M.
中科院分区：
文献类型：
--
作者：
Baltich, Jennifer;Hatch-Vallier, Leah;Adams, Aaron M.;Arruda, Ellen M.;Larkin, Lisa M.
通常需要神经移植物来替换因疾病、手术或大面积创伤而受损的组织。诸如移植物可用性、供体部位发病率和免疫排斥等限制使得研究人员开发了工程神经组织的策略。本研究的目的是使用胎儿神经细胞与成纤维细胞单层的共培养物制造无支架三维（3D）神经构建体，并允许共培养物重塑成具有外部成纤维细胞层和互连神经元细胞的内部核心的3D构建体。将原代成纤维细胞接种在层粘连蛋白包被的平板上，并使其形成融合的单层。将从E-15脊髓分离的神经细胞接种在成纤维细胞单层的顶部，并使其在成纤维细胞单层上形成网状单层。培养基移动引发成纤维细胞单层的收缩，并将共培养物重塑为通过两个限制销静态固定在适当位置的3D结构。使用S100（雪旺细胞）、β3-微管蛋白、DAPI和胶原蛋白I的免疫组织化学表明神经细胞的内核被成纤维细胞的外层包围。在体外测量3D神经和对照（仅成纤维细胞）结构的传导速度，并与新生儿坐骨神经的体内测量进行比较。神经结构的传导速度与24天的新生儿神经相当。雪旺细胞的存在和在体外传导神经信号的能力表明无支架3D神经构建体将是神经修复的可行选择。
Nerve grafts are often required to replace tissue damaged by disease, surgery, or extensive trauma. Limitations such as graft availability, donor site morbidity, and immune rejection have led investigators to develop strategies to engineer nerve tissue. The goal of this study was to fabricate a scaffoldless three-dimensional (3D) nerve construct using a co-culture of fetal nerve cells with a fibroblast monolayer and allow the co-culture to remodel into a 3D construct with an external fibroblast layer and an internal core of interconnected neuronal cells. Primary fibroblasts were seeded on laminin-coated plates and allowed to form a confluent monolayer. Neural cells isolated from E-15 spinal cords were seeded on top of the fibroblast monolayer and allowed to form a networked monolayer across the monolayer of fibroblasts. Media shifts initiated contraction of the fibroblast monolayer and a remodeling of the co-culture into a 3D construct held statically in place by the two constraint pins. Immunohisto-chemistry using S100 (Schwann cell), β3-tubulin, DAPI, and collagen I indicated an inner core of nerve cells surrounded by an external layer of fibroblasts. Conduction velocities of the 3D nerve and control (fibroblast-only) constructs were measured in vitro and compared to in vivo measures of neonatal sciatic nerve. The conduction velocities of the nerve constructs were comparable to 24-d-old neonatal nerve. The presence of Schwann cells and the ability to conduct neuronal signals in vitro suggest the scaffoldless 3D nerve constructs will be a viable option for nerve repair.
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影响因子：
41.2
作者：
Luo, Y;Shoichet, MS
通讯作者：
Shoichet, MS
影响因子：
--
作者：
Larkin, Lisa M.;Calve, Sarah;Arruda, Ellen M.
通讯作者：
Arruda, Ellen M.
影响因子：
2.1
作者：
Karagoz, Huseyin;Ulkur, Ersin;Celikoz, Bahattin
通讯作者：
Celikoz, Bahattin
影响因子：
3.5
作者：
MILLER, IJ;REEDY, FE
通讯作者：
REEDY, FE
影响因子：
14
作者：
Spilker, MH;Asano, K;Spector, M
通讯作者：
Spector, M