Dynamics of Intrinsic Glucose Uptake Kinetics in Human Mesenchymal Stem Cells During Chondrogenesis
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Chondrogenesis of human mesenchymal stem cells (hMSCs) is an important biological process in many applications including cartilage tissue engineering. We investigated the glucose uptake characteristics of aggregates of hMSCs undergoing chondrogenesis over a 3-week period both experimentally and by using a mathematical model. Initial concentrations of glucose in the medium were varied from 1 to 4.5 g/L to mimic limiting conditions and glucose uptake profiles were obtained. A reaction–diffusion mathematical model was implemented and solved to estimate kinetic parameters. Experimental glucose uptake rates increased with culture time for aggregates treated with higher initial glucose concentrations (3 and 4.5 g/L), whereas they decreased or remained constant for those treated with lower initial glucose concentrations (1 and 2 g/L). Lactate production rate increased by as much as 40% for aggregates treated with higher initial glucose concentrations (2, 3 and 4.5 g/L), whereas it remained constant for those treated with 1 g/L initial glucose concentration. The estimated DNA-normalized maximum glucose uptake rate decreased by a factor of 9 from day 0–2 (12.5 mmol/s/g DNA) to day 6–8 (1.5 mmol/s/g DNA), after which it started to increase. On day 18–20, its value (17.5 mmol/s/g DNA) was about 11 times greater than its lowest value. Further, the extracellular matrix levels of aggregates at day 14 and day 21 correlated with their overall glucose uptake and lactate production. The results suggest that during chondrogenesis, for optimal results, cells require increasing amounts of glucose. Our results also suggest that diffusion limitations play an important role in glucose uptake even in the smaller size aggregate model of chondrogenesis. Further, the results indicate that glucose uptake or lactate production can be a tool for predicting the end quality of tissue during the process of chondrogenesis. The estimated kinetic parameters can be used to model glucose requirements in cartilage tissue engineering applications.
KeywordsCartilage Aggregate cultures Mass transport Glucose consumption Diffusional limitation Michaelis–Menten kinetics Lactate production Mathematical modeling
The authors would like to thank Amad Awadallah and Dr. Don Lennon at the Skeletal Research Center, Department of Biology, Case Western Reserve University for help with histology and hMSC isolation respectively. This work is supported in part by a grant from the National Institutes of Health (EB021911). Dr. Motavalli is supported by a fellowship from the Case School of Engineering, College of Arts and Sciences and School of Medicine, Case Western Reserve University.
Conflict of interest
The authors declare no conflicts of interests related to the work performed in the manuscript.
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