1.1 本规程涵盖离心细胞粘附试验，可用于检测传代或处理后细胞粘附特性的变化。该方法测量将细胞从基质上分离所需的力。在许多变量中，粘附可能因细胞表型的变化而变化。 1.2 本规程不包括验证表面涂层均匀性的方法，也不包括表征表面的方法。 1.3 这些细胞可能包括来自任何物种的成体细胞、祖细胞或干细胞。细胞类型可包括软骨细胞、成纤维细胞、成骨细胞、胰岛细胞或其他相关粘附细胞类型。 1.4 本规程不包括分离或收获细胞的方法。本规程不包括通过免疫染色确定的定量基因表达变化或生物标记物类型或浓度变化的方法。 该实践也不包括定量图像分析技术。 1.5 本标准并非旨在解决与其使用相关的所有安全问题（如有）。本标准的用户有责任在使用前制定适当的安全、健康和环境实践，并确定监管限制的适用性。 1.6 本国际标准是根据世界贸易组织技术性贸易壁垒（TBT）委员会发布的《关于制定国际标准、指南和建议的原则的决定》中确立的国际公认标准化原则制定的。 ====意义和用途====== 4.1 本规程描述了一种细胞粘附方法，该方法可用于在给定的RCF下为粘附在基质上（通常为短时间）的细胞提供分离百分比。 当与同一基板上的参考（粘附）细胞类型进行比较时，通过本规程产生的信息可用于获得细胞与未涂覆或预涂覆基板粘附的半定量测量。如Reyes和Garcia（2003）所述，建议将50%点用于配体浓度或RCF，以最稳健地测量粘附强度。粘附可能因细胞表型的变化或表面的特定特性而变化。基质可包括经组织培养处理的聚苯乙烯、生物材料或生物活性表面。如果基质是水凝胶，则必须注意避免水凝胶中的内聚破坏（即，分离的细胞拉走了凝胶碎片）。涂层可能包括（但不限于）以下内容: 天然或合成生物材料、水凝胶、细胞外基质成分、配体、粘附或生物活性分子、基因或基因产物。细胞浓度也很关键，因为使用浓度过高的细胞可能会导致细胞作为薄片而不是单个细胞分离。这种离心方法一旦得到验证，可能适用于质量控制（QC）和产品开发。然而，在该方法与其他细胞粘附测量相关之前，当前方法应与其他已知的细胞粘附测量平行运行。 4.2 本规程不包括通过免疫染色确定的定量基因表达变化或生物标记物变化的方法。此外，本规程不包括定量图像分析技术。在某些情况下，粘合性能的变化可能会反映出差异化或破坏的程度- 细胞分化。然而，值得注意的是，粘附相互作用不一定反映特定细胞类型的分化状态，尽管在许多情况下确实如此。（参见 X1.3 用于软骨细胞的粘附。）

1.1 This practice covers a centrifugation cell adhesion assay that can be used to detect changes in adhesive characteristics of cells with passage or treatments. This approach measures the force required to detach cells from a substrate. Adhesion, among many variables, may vary due to changes in the phenotype of the cells. 1.2 This practice does not cover methods to verify the uniformity of coating of surfaces, nor does it cover methods for characterizing surfaces. 1.3 The cells may include adult, progenitor, or stem cells from any species. The types of cells may include chondrocytes, fibroblasts, osteoblast, islet cells, or other relevant adherent cell types. 1.4 This practice does not cover methods for isolating or harvesting of cells. This practice does not cover methods to quantitate changes in gene expression, or changes in biomarker type or concentration, as identified by immunostaining. Nor does this practice cover quantitative image analysis techniques. 1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use. 1.6 This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles for the Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee. ====== Significance And Use ====== 4.1 This practice describes a cell adhesion method that can be used to provide a detachment percent at a given RCF for cells that have adhered to a substrate, typically for a short time. The information generated by this practice can be used to obtain a semi-quantitative measurement of the adhesion of cells to either an uncoated or pre-coated substrate, when compared to a reference (adherent) cell type on the same substrate. As described in Reyes and Garcia (2003), it is recommended that the 50 % point be used for either ligand concentration or RCF for the most robust measurement of adhesion strength. The adhesion may vary due to changes in the phenotype of the cells or as a result of the specific properties of the surface. The substrate may include tissue culture-treated polystyrene, biomaterials, or bioactive surfaces. If the substrate is a hydrogel, care must be taken to avoid cohesive failure in the hydrogel (that is, detached cells have pulled away fragments of gel). The coating may consist of (but is not limited to) the following: natural or synthetic biomaterials, hydrogels, components of extracellular matrix (ECM), ligands, adhesion or bioactive molecules, genes, or gene products. Cell concentration is also critical, as use of too high a concentration of cells may result in cells detaching as a sheet, rather than as individual cells. This centrifugation approach, once validated, may be applicable for quality control (QC) and product development. However, until the method is correlated to other measures of cell attachment, the current method should be run in parallel with other known measures of cell adhesion. 4.2 This practice does not cover methods to quantitate changes in gene expression, or changes in biomarkers, as identified by immunostaining. This practice additionally does not cover quantitative image analysis techniques. In some cases, the change in adhesive properties may reflect on the degree of differentiation or de-differentiation of the cells. However, it is worth noting that adhesive interactions do not necessarily reflect the differentiation state of a particular cell type, although in many instances they do. (See X1.3 for application to the adhesion of chondrocytes.)