Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA.
Biology Department, Miami University, Oxford, OH, USA.
Cartilage. 2021 Dec;13(2_suppl):95S-106S. doi: 10.1177/1947603520906605. Epub 2020 Feb 26.
Glucose concentrations used in current cell culture methods are a significant departure from physiological glucose levels. The study focuses on comparing the effects of glucose concentrations on primary human progenitors (connective tissue progenitors [CTPs]) used for cartilage repair.
Cartilage- (Outerbridge grade 1, 2, 3; superficial and deep zone cartilage), infrapatellar fatpad-, synovium-, and periosteum-derived cells were obtained from 63 patients undergoing total knee arthroplasty and cultured simultaneously in fresh chondrogenic media containing 25 mM glucose (HGL) or 5 mM glucose (NGL) for pairwise comparison. Automated ASTM-based quantitative image analysis was used to determine colony-forming efficiency (CFE), effective proliferation rates (EPR), and sulfated-proteoglycan (GAG-ECM) staining of the CTPs across tissue sources.
HGL resulted in increased cell cultures with CFE = 0 compared with NGL in all tissue sources ( = 0.049). The CFE in NGL was higher than HGL for superficial cartilage ( < 0.001), and contrary for synovium-derived CTPs ( = 0.046) when CFE > 0. EPR of the CTPs did not differ between the media in the 6-day assay time period ( = 0.082). The GAG-ECM area of the CTPs and their progeny was increased in presence of HGL ( = 0.027).
Glucose concentration is critical to progenitor's physiology and should be taken into account in the setting of protocols for clinical or cell expansion strategies.
目前细胞培养方法中使用的葡萄糖浓度与生理葡萄糖水平有很大的差异。本研究旨在比较葡萄糖浓度对用于软骨修复的原代人祖细胞（结缔组织祖细胞[CTP]）的影响。
从 63 例行全膝关节置换术的患者中获取软骨（Outerbridge 分级 1、2、3；浅部和深部软骨）、髌下脂肪垫、滑膜和骨膜来源的细胞，并同时在含有 25mM 葡萄糖（HGL）或 5mM 葡萄糖（NGL）的新鲜软骨形成培养基中进行培养，进行成对比较。采用基于 ASTM 的自动定量图像分析来确定 CTP 的集落形成效率（CFE）、有效增殖率（EPR）和硫酸软骨素-ECM 染色。
与 NGL 相比，HGL 导致所有组织来源的 CFE = 0 的细胞培养增加（= 0.049）。在 NGL 中，CFE 高于 HGL 的组织来源为浅部软骨（<0.001），而对于滑膜来源的 CTPs 则相反（= 0.046），当 CFE > 0 时。在 6 天的检测时间内，两种培养基中 CTPs 的 EPR 没有差异（= 0.082）。在 HGL 存在的情况下，CTP 及其后代的 GAG-ECM 面积增加（= 0.027）。
葡萄糖浓度对祖细胞的生理学至关重要，在临床或细胞扩增策略的方案中应考虑到这一点。
