Comparative study of expression and activity of glucose transporters between stem cell-derived brain microvascular endothelial cells and hCMEC/D3 cells

Abstract

Glucose constitutes a major source of energy of mammalian brains. Glucose uptake at the blood-brain barrier (BBB) occurs through a facilitated glucose transport, through glucose transporter 1 (GLUT1), although other isoforms have been described at the BBB. Mutations in GLUT1 are associated with the GLUT1 deficiency syndrome, yet none of the current in vitro models of the human BBB maybe suited for modeling such a disorder. In this study, we investigated the expression of glucose transporters and glucose diffusion across brain microvascular endothelial cells (BMECs) derived from healthy patient-derived induced pluripotent stem cells (iPSCs). We investigated the expression of different glucose transporters at the BBB using immunocytochemistry and flow cytometry and measured glucose uptake and diffusion across BMEC monolayers obtained from two iPSC lines and from hCMEC/D3 cells. BMEC monolayers showed expression of several glucose transporters, in particular GLUT1, GLUT3, and GLUT4. Diffusion of glucose across the monolayers was mediated via a saturable transcellular mechanism and partially inhibited by pharmacological inhibitors. Taken together, our study suggests the presence of several glucose transporters isoforms at the human BBB and demonstrates the feasibility of modeling glucose across the BBB using patient-derived stem cells.

Neurolysin substrates bradykinin, neurotensin and substance P enhance brain microvascular permeability in a human in vitro model

“…In the present study, we assessed the direct effect of Nln substrates BK, NT and SP on the permeability of BBB in vitro using human stem cell‐derived, brain microvascular endothelial cells (iPSC‐derived BMECs) in culture. Our observations indicate that all three peptides enhance BBB permeability in iPSC‐derived BMECs similar to that of hCMEC/D3 cells (Figure 1), a widely used human brain microvascular endothelial cell line 27,33‐41 . The observed effects of BK, NT and SP on the permeability of BBB in iPSC‐derived BMECs were concentration‐dependent, showing a pronounced effect at 1 µmol L ‐1 and marginal or no effects at concentrations of 0.1 and 0.5 µmol L ‐1 (Figure 2).…”

The S1 protein of SARS-CoV-2 crosses the blood–brain barrier in mice

“…For that reason, we used in vitro models of the human BBB, which can be useful in studying mechanisms of BBB permeability. The model used in this study is derived from human iPSCs and develops a brain endothelial cell-like phenotype that includes functional BBB influx and efflux transporters and strong barrier properties that permit the study of transport without confounding effects of high baseline leakage 46,47 . In this model, we did not observe significant differences in permeability for I-S1 compared to T-Alb.…”

Establishment of an in Vitro Human Blood-Brain Barrier Model Derived from Induced Pluripotent Stem Cells and Comparison to a Porcine Cell-Based System

“…The hiPSC-derived BMECs expressed the efflux transporters PGP and BCRP ( Figure 3A (Cl1)), which play a critical role at the BBB to prevent the entry of harmful molecules into the brain [25]. Members of the solute carrier family transporters (SLCs) such as GLUT-1 (SLC2A1), the major glucose transporter at the BBB and GLUT-3 (SLC2A3) were also expressed [26,27] (Figure 3A (Cl1) and Figure S2 (Cl2)). To better characterize the molecular nature of hiPSC-derived BMECs in co-culture with human astrocytes (at d1 co-culture), we performed whole genome expression analysis by RNA sequencing.…”