A Hydrogel Model of the Human Blood-brain Barrier Using Differentiated Stem Cells

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2023 Apr 4

PMID 37014916

Authors

Nandita Rahatekar Singh

Radka Gromnicova

Andreas Brachner

Igor Kraev

Ignacio A Romero

Winfried Neuhaus

David Male

Affiliations

Soon will be listed here.

Abstract

An in vitro model of the human blood-brain barrier was developed, based on a collagen hydrogel containing astrocytes, overlaid with a monolayer of endothelium, differentiated from human induced pluripotent stem cells (hiPSCs). The model was set up in transwell filters allowing sampling from apical and basal compartments. The endothelial monolayer had transendothelial electrical resistance (TEER) values >700Ω.cm2 and expressed tight-junction markers, including claudin-5. After differentiation of hiPSCs the endothelial-like cells expressed VE-cadherin (CDH5) and von-Willebrand factor (VWF) as determined by immunofluorescence. However, electron microscopy indicated that at set-up (day 8 of differentiation), the endothelial-like cells still retained some features of the stem cells, and appeared immature, in comparison with primary brain endothelium or brain endothelium in vivo. Monitoring showed that the TEER declined gradually over 10 days, and transport studies were best carried out in a time window 24-72hrs after establishment of the model. Transport studies indicated low permeability to paracellular tracers and functional activity of P-glycoprotein (ABCB1) and active transcytosis of polypeptides via the transferrin receptor (TFR1).

Citing Articles

Blood-Brain Barrier-Targeting Nanoparticles: Biomaterial Properties and Biomedical Applications in Translational Neuroscience.

Asimakidou E, Tan J, Zeng J, Lo C Pharmaceuticals (Basel). 2024; 17(5).

PMID: 38794182 PMC: 11123901. DOI: 10.3390/ph17050612.

Characterization of an iPSC-based barrier model for blood-brain barrier investigations using the SBAD0201 stem cell line.

Ozgur B, Puris E, Brachner A, Appelt-Menzel A, Oerter S, Balzer V Fluids Barriers CNS. 2023; 20(1):96.

PMID: 38115090 PMC: 10731806. DOI: 10.1186/s12987-023-00501-9.

Subcellular trafficking and transcytosis efficacy of different receptor types for therapeutic antibody delivery at the blood‒brain barrier.

Holst M, de Wit N, Ozgur B, Brachner A, Hyldig K, Appelt-Menzel A Fluids Barriers CNS. 2023; 20(1):82.

PMID: 37932749 PMC: 10626680. DOI: 10.1186/s12987-023-00480-x.

Blood-Brain Barrier Breakdown in Neuroinflammation: Current In Vitro Models.

Brandl S, Reindl M Int J Mol Sci. 2023; 24(16).

PMID: 37628879 PMC: 10454051. DOI: 10.3390/ijms241612699.

Experimental Models to Study the Functions of the Blood-Brain Barrier.

Lach A, Wnuk A, Wojtowicz A Bioengineering (Basel). 2023; 10(5).

PMID: 37237588 PMC: 10215847. DOI: 10.3390/bioengineering10050519.

References

Lajoie J, Shusta E . Targeting receptor-mediated transport for delivery of biologics across the blood-brain barrier. Annu Rev Pharmacol Toxicol. 2014; 55:613-31. PMC: 5051266. DOI: 10.1146/annurev-pharmtox-010814-124852. View

Grifno G, Farrell A, Linville R, Arevalo D, Kim J, Gu L . Tissue-engineered blood-brain barrier models via directed differentiation of human induced pluripotent stem cells. Sci Rep. 2019; 9(1):13957. PMC: 6764995. DOI: 10.1038/s41598-019-50193-1. View

Warren M, Zerangue N, Woodford K, Roberts L, Tate E, Feng B . Comparative gene expression profiles of ABC transporters in brain microvessel endothelial cells and brain in five species including human. Pharmacol Res. 2009; 59(6):404-13. DOI: 10.1016/j.phrs.2009.02.007. View

Katt M, Linville R, Mayo L, Xu Z, Searson P . Functional brain-specific microvessels from iPSC-derived human brain microvascular endothelial cells: the role of matrix composition on monolayer formation. Fluids Barriers CNS. 2018; 15(1):7. PMC: 5819713. DOI: 10.1186/s12987-018-0092-7. View

Kurosawa T, Tega Y, Higuchi K, Yamaguchi T, Nakakura T, Mochizuki T . Expression and Functional Characterization of Drug Transporters in Brain Microvascular Endothelial Cells Derived from Human Induced Pluripotent Stem Cells. Mol Pharm. 2018; 15(12):5546-5555. DOI: 10.1021/acs.molpharmaceut.8b00697. View

Demeule M, Currie J, Bertrand Y, Che C, Nguyen T, Regina A . Involvement of the low-density lipoprotein receptor-related protein in the transcytosis of the brain delivery vector angiopep-2. J Neurochem. 2008; 106(4):1534-44. DOI: 10.1111/j.1471-4159.2008.05492.x. View

Canfield S, Stebbins M, Faubion M, Gastfriend B, Palecek S, Shusta E . An isogenic neurovascular unit model comprised of human induced pluripotent stem cell-derived brain microvascular endothelial cells, pericytes, astrocytes, and neurons. Fluids Barriers CNS. 2019; 16(1):25. PMC: 6685239. DOI: 10.1186/s12987-019-0145-6. View

Veszelka S, Toth A, Walter F, Toth A, Grof I, Meszaros M . Comparison of a Rat Primary Cell-Based Blood-Brain Barrier Model With Epithelial and Brain Endothelial Cell Lines: Gene Expression and Drug Transport. Front Mol Neurosci. 2018; 11:166. PMC: 5972182. DOI: 10.3389/fnmol.2018.00166. View

Zhang W, Liu Q, Haqqani A, Leclerc S, Liu Z, Fauteux F . Differential expression of receptors mediating receptor-mediated transcytosis (RMT) in brain microvessels, brain parenchyma and peripheral tissues of the mouse and the human. Fluids Barriers CNS. 2020; 17(1):47. PMC: 7376922. DOI: 10.1186/s12987-020-00209-0. View

10.

Butt A . Effect of inflammatory agents on electrical resistance across the blood-brain barrier in pial microvessels of anaesthetized rats. Brain Res. 1995; 696(1-2):145-50. DOI: 10.1016/0006-8993(95)00811-4. View

11.

Weksler B, Romero I, Couraud P . The hCMEC/D3 cell line as a model of the human blood brain barrier. Fluids Barriers CNS. 2013; 10(1):16. PMC: 3623852. DOI: 10.1186/2045-8118-10-16. View

12.

Appelt-Menzel A, Cubukova A, Gunther K, Edenhofer F, Piontek J, Krause G . Establishment of a Human Blood-Brain Barrier Co-culture Model Mimicking the Neurovascular Unit Using Induced Pluri- and Multipotent Stem Cells. Stem Cell Reports. 2017; 8(4):894-906. PMC: 5390136. DOI: 10.1016/j.stemcr.2017.02.021. View

13.

Lippmann E, Al-Ahmad A, Azarin S, Palecek S, Shusta E . A retinoic acid-enhanced, multicellular human blood-brain barrier model derived from stem cell sources. Sci Rep. 2014; 4:4160. PMC: 3932448. DOI: 10.1038/srep04160. View

14.

Paterson J, Webster C . Exploiting transferrin receptor for delivering drugs across the blood-brain barrier. Drug Discov Today Technol. 2016; 20:49-52. DOI: 10.1016/j.ddtec.2016.07.009. View

15.

Daneman R, Zhou L, Agalliu D, Cahoy J, Kaushal A, Barres B . The mouse blood-brain barrier transcriptome: a new resource for understanding the development and function of brain endothelial cells. PLoS One. 2010; 5(10):e13741. PMC: 2966423. DOI: 10.1371/journal.pone.0013741. View

16.

Wilhelm I, Krizbai I . In vitro models of the blood-brain barrier for the study of drug delivery to the brain. Mol Pharm. 2014; 11(7):1949-63. DOI: 10.1021/mp500046f. View

17.

Kamiichi A, Furihata T, Kishida S, Ohta Y, Saito K, Kawamatsu S . Establishment of a new conditionally immortalized cell line from human brain microvascular endothelial cells: a promising tool for human blood-brain barrier studies. Brain Res. 2012; 1488:113-22. DOI: 10.1016/j.brainres.2012.09.042. View

18.

Pulous F, Petrich B . Integrin-dependent regulation of the endothelial barrier. Tissue Barriers. 2019; 7(4):1685844. PMC: 6866814. DOI: 10.1080/21688370.2019.1685844. View

19.

Rowland T, Miller L, Blaschke A, Doss E, Bonham A, Hikita S . Roles of integrins in human induced pluripotent stem cell growth on Matrigel and vitronectin. Stem Cells Dev. 2009; 19(8):1231-40. DOI: 10.1089/scd.2009.0328. View

20.

Wilson H, Faubion M, Hjortness M, Palecek S, Shusta E . Cryopreservation of Brain Endothelial Cells Derived from Human Induced Pluripotent Stem Cells Is Enhanced by Rho-Associated Coiled Coil-Containing Kinase Inhibition. Tissue Eng Part C Methods. 2016; 22(12):1085-1094. PMC: 5175444. DOI: 10.1089/ten.TEC.2016.0345. View