Liu Jennifer W, Lin Kevin H, Weber Christian, Bhalla Sameer, Kelso Sean, Wang Kaixi, Tang Simon Y
Department of Biomedical Engineering, Washington University in St. Louis; Department of Orthopaedic Surgery, Washington University in St. Louis.
Department of Biology, Washington University in St. Louis.
J Vis Exp. 2017 Apr 11(122):55437. doi: 10.3791/55437.
Intervertebral disc (IVD) degeneration is a significant contributor to low back pain. The IVD is a fibrocartilaginous joint that serves to transmit and dampen loads in the spine. The IVD consists of a proteoglycan-rich nucleus pulposus (NP) and a collagen-rich annulus fibrosis (AF) sandwiched by cartilaginous end-plates. Together with the adjacent vertebrae, the vertebrae-IVD structure forms a functional spine unit (FSU). These microstructures contain unique cell types as well as unique extracellular matrices. Whole organ culture of the FSU preserves the native extracellular matrix, cell differentiation phenotypes, and cellular-matrix interactions. Thus, organ culture techniques are particularly useful for investigating the complex biological mechanisms of the IVD. Here, we describe a high-throughput approach for culturing whole lumbar mouse FSUs that provides an ideal platform for studying disease mechanisms and therapies for the IVD. Furthermore, we describe several applications that utilize this organ culture method to conduct further studies including contrast-enhanced microCT imaging and three-dimensional high-resolution finite element modeling of the IVD.
椎间盘（IVD）退变是导致腰痛的一个重要因素。IVD是一个纤维软骨关节，用于传递和缓冲脊柱中的负荷。IVD由富含蛋白聚糖的髓核（NP）和富含胶原蛋白的纤维环（AF）组成，两者被软骨终板夹在中间。与相邻椎体一起，椎体-椎间盘结构形成一个功能脊柱单元（FSU）。这些微观结构包含独特的细胞类型以及独特的细胞外基质。FSU的全器官培养可保留天然细胞外基质、细胞分化表型以及细胞与基质的相互作用。因此，器官培养技术对于研究IVD复杂的生物学机制特别有用。在此，我们描述一种用于培养整个腰椎小鼠FSU的高通量方法，该方法为研究IVD的疾病机制和治疗方法提供了一个理想平台。此外，我们描述了几种利用这种器官培养方法进行进一步研究的应用，包括对比增强微型计算机断层扫描（microCT）成像和IVD的三维高分辨率有限元建模。
