Amino Acid Uptake Limitations during Human Mesenchymal Stem Cell-Based Chondrogenesis

Abstract

A mino acids are the essential building blocks for collagen and proteoglycan, which are the main constituents for cartilage extracellular matrix (ECM). Synthesis of ECM proteins requires the uptake of various essential/nonessential amino acids. Analyzing amino acid metabolism during chondrogenesis can help to relate tissue quality to amino acid metabolism under different conditions. In our study, we studied amino acid uptake/secretion using human mesenchymal stem cell (hMSC)-based aggregate chondrogenesis in a serum-free induction medium with a defined chemical formulation. The initial glucose level and medium-change frequency were varied. Our results showed that essential amino acid uptake increased with time during hMSCs chondrogenesis for all initial glucose levels and medium-change frequencies. Essential amino acid uptake rates were initial glucose-level independent. The DNA-normalized glycosaminoglycans and hydroxyproline content of chondrogenic aggregates correlated with cumulative uptake of leucine, valine, and tryptophan regardless of initial glucose levels and medium-change frequencies. Collectively, our results show that amino acid uptake rates during in vitro chondrogenesis were insufficient to produce a tissue with an ECM content similar to that of human neonatal cartilage or adult cartilage. Furthermore, this deficiency was likely related to the downregulation of some key amino acid transporters in the cells. Such deficiency could be partially improved by increasing the amino acid availability in the chondrogenic medium by changing culture conditions.

Impact Statement

Human mesenchymal stem cells (hMSCs) are a promising cell source for cartilage tissue engineering (TE). However, the state-of-the-art cartilage TE still falls short of clinical expectations as the engineered tissue differs from native hyaline cartilage, especially in extracellular matrix (ECM) content and composition. Amino acids are the building blocks for proteins and are critical for cartilage ECM synthesis. Despite their importance, to our knowledge, no studies investigated amino acid uptake during hMSCs chondrogenesis. This study reports amino acid uptake during in vitro chondrogenesis using hMSCs aggregate system.