Optimal Hypoxic Preconditioning of Human Embryonic Stem Cell-Derived Mesenchymal Stem Cells (hES-MSCs) and Their Characteristics

Abstract

Background and Objectives: Hypoxia is frequently used to enhance stem cell function. However, the optimal level of hypoxia for growth and function of human embryonic stem cell-derived mesenchymal stem cells (hES-MSCs) is yet to be determined. The purpose of this study was to find the optimal level of hypoxia for hES-MSCs and characteristics of hES-MSCs cultured under these optimal hypoxic conditions. Methods and Results: Cell viability and changes in the morphology of hES-MSCs were determined through cell proliferation and CCK-8 assay. The hES-MSCs were preconditioned under various hypoxic conditions (0.5∼5% O2 and 24∼72 h). The expression of cytokines in each culture condition was compared using cytokine array analysis. The morphology of hES-MSCs did not change under various hypoxic culture conditions. hES-MSCs viability after 48 h incubation in 2% O2 condition was higher than that in normoxic condition. HIF1α expression was increased up to six folds after 48 h of hypoxic preconditioning. HIF1α expression in hES-MSCs peaked after 48 h of incubation in 1% O2 condition. The expressions of PDGF-BB, IGFBP-6, VEGF-A, and angiogenin were increased after hES-MSCs were incubated for 48 h in 2% O2 condition. Conclusions: The hES-MSCs viability and expressions of PDGF-BB, IGFBP-6, VEGF-A, and angiogenin increased after 48 h incubation in 2% O2 condition.

Role of hypoxia preconditioning in therapeutic potential of mesenchymal stem-cell-derived extracellular vesicles

“…Moderate (< 5% to > 2% O 2 ), severe (≤ 2% to ≥ 0.1% O 2 ), and anoxia (< 0.1% O 2 ) are hypoxia levels equal or below 5% oxygen concentration. Depending on O 2 concentration and hypoxia time, cells show different responses, as observed in human embryonic-derived MSC[ 42 ]. That occurs mainly through activation of hypoxia-inducible factor (HIF).…”

The impact of hypoxia preconditioning on mesenchymal stem cells performance in hypertensive kidney disease

“…Overstretched cells are perhaps not optimal for exerting desirable paracrine effects. The survival rate of MSCs decreases even further at 72 h with 1% O 2 as compared to 24 and 48 h suggesting that excessive HIF-1α expression negatively affects cell survival [ 53 ].…”

Decellularized extracellular matrix-based scaffold and hypoxic priming: A promising combination to improve the phenotype of degenerate intervertebral disc cells

“…1. In order to recreate the unique microenvironment defined by low oxygen in which cells reside [18][19][20], IVD cells were initially grown in 2D monolayers under hypoxia (2% pO 2 ) for 48 h which is an adequate period for several experimental models to perform hypoxic preconditioning [37,38]. The behavior of hypoxic preconditioned IVD cells was compared with that of the same cells cultured under normoxia (21% pO 2 ).…”