Elsevier

Medical Engineering & Physics

Volume 48, October 2017, Pages 3-13
Medical Engineering & Physics

Engineering human renal epithelial cells for transplantation in regenerative medicine

https://doi.org/10.1016/j.medengphy.2017.03.009Get rights and content
Under a Creative Commons license
open access

Highlights

  • 3D culture of hRECs preserves long-term cell viability and trophic factor secretion.

  • Genetically engineered hRECs retain long-term cell viability and secretome.

  • Conformal coating of hRECs improves long-term cell viability and secretome.

Abstract

Cellular transplantation may treat several human diseases by replacing damaged cells and/or providing a local source of trophic factors promoting regeneration. We utilized human renal epithelial cells (hRECs) isolated from cadaveric donors as a cell model. For efficacious implementation of hRECs for treatment of kidney diseases, we evaluated a novel encapsulation strategy for immunoisolation of hRECs and lentiviral transduction of the Green Fluorescent Protein (GFP) as model gene for genetic engineering of hRECs to secrete desired trophic factors. In specific, we determined whether encapsulation through conformal coating and/or GFP transduction of hRECs allowed preservation of cell viability and of their trophic factor secretion. To that end, we optimized cultures of hRECs and showed that aggregation in three-dimensional spheroids significantly preserved cell viability, proliferation, and trophic factor secretion. We also showed that both wild type and GFP-engineered hRECs could be efficiently encapsulated within conformal hydrogel coatings through our fluid dynamic platform and that this resulted in further improvement of cell viability and trophic factors secretion. Our findings may lay the groundwork for future therapeutics based on transplantation of genetically engineered human primary cells for treatment of diseases affecting kidneys and potentially other tissues.

Keywords

Three-dimensional culture
Cell encapsulation
Conformal coating
Hydrogel
Gene therapy
Trophic factors

Abbreviations

2D
Two-dimensional
3D
Three-dimensional
ALG
alginate
CCE
conformal coating encapsulation
DTT
dithiothreitol
dVS
divinyl sulfone
EGF
epidermal growth factor
FGFb
basic fibroblast growth factor
GFP
green fluorescent protein
GM-CSF
granulocyte macrophage colony-stimulating factor
HBSS
Hanks' balanced salt solution
HGF
hepatocyte growth factor
hRECs
Human Renal Epithelial Cells
MCP-1
monocyte chemotactic protein-1 (CCL2)
MMP-2
matrix metalloproteinases-2
MOI
multiplicity of infection
MTT
3-(4,5-Dimethylthiazol-2-yl)−2,5-diphenyltetrazolium bromide
PAI-1
plasminogen activator inhibitor-1
PBS
phosphate-buffered saline
PEG
polyethylene glycol
pHEMA
Poly(2-hydroxyethyl methacrylate)
PPG
polypropylene glycol
RPM
revolutions per minute
SCF
stem cell factor
TEA
triethanolamine
TGFα
transforming growth factor alpha
TIMP
tissue inhibitor of metalloproteinase
UP-MVG
ultrapure medium viscosity (> 200 mPas) sodium alginate where minimum 60% of the monomer units are guluronate
VCAM-1
vascular cell adhesion molecule-1
VEGF
vascular endothelial growth factor

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