Andrade Anenisia C, Chrysis Dionisios, Audi Laura, Nilsson Ola
Endocr Dev. 2011;21:52-66. doi: 10.1159/000328127. Epub 2011 Aug 22.
In humans, the study of linear growth at the level of growth plate is limited to the use of mainly noninvasive methods. The use of in vitro studies and diverse animal models are alternative applied methods to unravel the underlying mechanisms of endochondral ossification and bone remodeling. In this chapter, we will outline important techniques to study molecular interactions and the regulation of gene expression in chondrocytes as well as bone cells. Microdissection of growth plate provides a powerful approach to study gene expression of individual zones and temporal senescence of growth plate cartilage by using microarray and real-time PCR analysis. IHC gives valuable information about the distribution of proteins of interest throughout distinct layers of the growth plate. The TUNEL assay has been successfully applied to study apoptosis in chondrocytes. In addition, organ(metatarsal) culture, primary and cell line cultures have been widely used to explore the regulation of chondrocyte differentiation under varying experimental conditions. Ultimately, these methods are essential to give new insights into the search for new diagnostic and therapeutic approaches for growth and various metabolic bone disorders.
在人类中，对生长板水平线性生长的研究主要限于使用非侵入性方法。体外研究和各种动物模型的应用是揭示软骨内骨化和骨重塑潜在机制的替代方法。在本章中，我们将概述研究软骨细胞以及骨细胞中分子相互作用和基因表达调控的重要技术。生长板的显微切割通过使用微阵列和实时PCR分析，为研究生长板软骨各个区域的基因表达和时间性衰老提供了一种强大的方法。免疫组织化学（IHC）提供了有关目标蛋白在生长板不同层中的分布的有价值信息。TUNEL检测已成功应用于研究软骨细胞中的凋亡。此外，器官（跖骨）培养、原代培养和细胞系培养已被广泛用于探索不同实验条件下软骨细胞分化的调控。最终，这些方法对于深入寻找生长和各种代谢性骨疾病的新诊断和治疗方法至关重要。
