Human Elastic Cartilage Engineering from Cartilage Progenitor Cells Using Rotating Wall Vessel Bioreactor
Human Elastic Cartilage Engineering from Cartilage Progenitor Cells Using Rotating Wall Vessel Bioreactor
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DOI：
10.1016/j.transproceed.2012.03.038
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发表时间：
2012-05-01
影响因子：
0.9
通讯作者：
Taniguchi, H.
中科院分区：
文献类型：
--
作者：
Takebe, T.;Kobayashi, S.;Taniguchi, H.
生物工程弹性软骨移植被认为是治疗颅面部缺损的一种很有前途的方法。我们先前已经证明，人耳软骨膜中含有大量的软骨前体细胞(CPC)。本研究的目的是研究使用旋转壁容器(RWV)生物反应器在体外构建三维弹性软骨。从人耳软骨膜分离出CPC，在二维培养条件下扩增并分化为软骨细胞。将完全分化的CPC种植到最近开发的PC-HAP/CHS(由胶原、羟基磷灰石和硫化软骨组成的多孔材料)支架上，并利用RWV生物反应器进行三维培养。3D工程构建物看起来闪闪发光，具有淡黄色、软骨样的形态。经RWV培养后，成型支架的形状保持不变。苏木精-伊红染色显示PC-HAP/CHS内有CPC的植入。Alcian蓝和Elastica Van Gieson染色显示有蛋白多糖和弹性纤维，它们是弹性软骨特有的细胞外基质。因此，在RWV生物反应器中进行三维培养后，人CPC形成了弹性软骨样组织。这些技术可能有助于未来在体外重建由弹性软骨组成的复杂结构的努力。
Transplantation of bioengineered elastic cartilage is considered to be a promising approach for patients with craniofacial defects. We have previously shown that human ear perichondrium harbors a population of cartilage progenitor cells (CPCs). The aim of this study was to examine the use of a rotating wall vessel (RWV) bioreactor for CPCs to engineer 3-D elastic cartilage in vitro. Human CPCs isolated from ear perichondrium were expanded and differentiated into chondrocytes under 2-D culture conditions. Fully differentiated CPCs were seeded into recently developed pC-HAp/ChS (porous material consisted of collagen, hydroxyapatite, and chondroitinsulfate) scaffolds and 3-D cultivated utilizing a RWV bioreactor. 3-D engineered constructs appeared shiny with a yellowish, cartilage-like morphology. The shape of the molded scaffold was maintained after RWV cultivation. Hematoxylin and eosin staining showed engraftment of CPCs inside pC-HAp/ChS. Alcian blue and Elastica Van Gieson staining showed of proteoglycan and elastic fibers, which are unique extracellular matrices of elastic cartilage. Thus, human CPCs formed elastic cartilage-like tissue after 3-D cultivation in a RWV bioreactor. These techniques may assist future efforts to reconstruct complicate structures composed of elastic cartilage in vitro.