Influence of Glucose Concentration on Colony-Forming Efficiency and Biological Performance of Primary Human Tissue-Derived Progenitor Cells.

OBJECTIVE

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.

DESIGN

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.

RESULTS

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).

CONCLUSION

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.