EVALUATION OF FLUORESCEIN DIACETATE FOR FLOW CYTOMETRIC DETERMINATION OF CELL VIABILITY IN ORTHOPEDIC RESEARCH
EVALUATION OF FLUORESCEIN DIACETATE FOR FLOW CYTOMETRIC DETERMINATION OF CELL VIABILITY IN ORTHOPEDIC RESEARCH
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DOI：
10.1002/jor.1100060402
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发表时间：
1988-07-01
影响因子：
2.8
通讯作者：
MANKIN, HJ
中科院分区：
文献类型：
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作者：
BELL, RS;BOURRET, LA;MANKIN, HJ
准确估计细胞活力在研究和骨科临床实践方面都是重要的。我们一直感兴趣的荧光素二乙酸酯（FDA）在评估软骨细胞存活冷冻保存后的骨软骨同种异体移植物，以及在评估肉瘤坏死术前化疗的潜力流式细胞术的应用。为了评估该方法用于细胞活力测定的适用性，本研究将FDA与更传统的方法（台盼蓝、克隆形成测定、代谢活性分析、DNA合成测量和坏死组织学评估）进行了比较。软骨细胞和肉瘤细胞在活力分析之前暴露于各种实验损伤。尽管从这些实验中可以明显看出，FDA准确地反映了物理损伤后的细胞存活，但它低估了化疗对体外细胞繁殖潜力的影响。然而，FDA与体内化疗后肿瘤活力的组织学评估高度相关。显然，选择用于测定活力的方法应该适合于实验损伤的类型，并且应该分析细胞功能（即，代谢活性或生殖能力）。
Accurate estimation of cellular viability is important both in research and in aspects of orthopaedic clinical practice. We have been interested in the potential for flow cytometric application of fluorescein diacetate (FDA) in evaluating chondrocyte survival following cryopreservation of osteochondral allografts as well as in the assessment of sarcoma necrosis following preoperative chemotherapy. In order to evaluate the suitability of this method for cell viability assays, this study compared FDA with more traditional methodology (trypan blue, clonigenic assay, metabolic activity analysis, measurement of DNA synthesis, and histological assessment of necrosis). Both chondrocytes and sarcoma cells were exposed to various experimental injuries prior to viability analysis. although it is is evident from these experiments that FDA accurately reflects cell survival after physical injury, it underestimates the effect of chemotherapy on cell reproductive potential in vitro. However, FDA is highly correlated with histological assessment of tumor viability after chemotherapy in vivo. It is apparent that the methodology chosen for determination of viability should be appropriate for the type of experimental injury and should analyze the cell function (i.e., metabolic activity or reproductive capacity) that is appropriate for the experimental model.