Vernon Robert B, Gooden Michel D
The Hope Heart Institute, Seattle, Washington 98104, USA.
In Vitro Cell Dev Biol Anim. 2002 Feb;38(2):97-101. doi: 10.1290/1071-2690(2002)038<0097:AIMFTC>2.0.CO;2.
The collagen gel contraction (CGC) assay is used frequently to study the cell-mediated reorganization of the extracellular natrix. In a typical CGC assay, cells embedded in a disk-shaped lattice (gel) of native type I collagen fibers compress the fibers and, consequently, reduce the diameter of the collagen disk within h or d. The degree to which the collagen is contracted is usually quantified by measurement of the diameter or the area of the disk. During CCC assays, friction or adhesion (or both) between gels and their culture containers can cause gels to be incompletely contracted or to acquire distorted shapes. Such occurrences degrade the reproducibility and reliability of measurements of gel dimensions. To address these problems, we developed an oil-supported collagen retraction (OSCR) assay that creates an environment of low friction and adhesion around the contracting collagen gel. The OSCR assay is accomplished with simple equipment and is easily performed, sensitive, and consistently yields fully contracted gels with minimal distortion.
胶原蛋白凝胶收缩(CGC)测定法常用于研究细胞介导的细胞外基质重组。在典型的CGC测定中,嵌入天然I型胶原纤维盘状晶格(凝胶)中的细胞压缩纤维,从而在数小时或数天内减小胶原盘的直径。胶原收缩的程度通常通过测量盘的直径或面积来量化。在CGC测定过程中,凝胶与其培养容器之间的摩擦或粘附(或两者兼有)会导致凝胶收缩不完全或形状扭曲。这些情况会降低凝胶尺寸测量的可重复性和可靠性。为了解决这些问题,我们开发了一种油支持的胶原回缩(OSCR)测定法,该方法在收缩的胶原凝胶周围创造了一个低摩擦和低粘附的环境。OSCR测定法使用简单的设备即可完成,操作简便、灵敏,并且始终能产生收缩完全且变形最小的凝胶。