Thermo-responsive cell culture carriers based on poly(Vinyl Methyl ether) - the effect of biomolecular ligands to balance cell adhesion and stimulated detachment.

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

Abstract

cTwo established material systems for thermally stimulated detachment of adherent cells were combined in a cross-linked polymer blend to merge favorable properties. Through this approach poly(N-isopropylacrylamide) (PNiPAAm) with its superior switching characteristic was paired with a poly(vinyl methyl ether)-based composition that allows adjusting physico-chemical and biomolecular properties in a wide range. Beyond pure PNiPAAm, the proposed thermoresponsive coating provides thickness, stiffness and swelling behavior, as well as an apposite density of reactive sites for biomolecular functionalization, as effective tuning parameters to meet specific requirements of a particular cell type regarding initial adhesion and ease of detachment. To illustrate the strength of this approach, the novel cell culture carrier was applied to generate transplantable sheets of human corneal endothelial cells (HCEC). Sheets were grown, detached, and transferred onto planar targets. Cell morphology, viability and functionality were analyzed by immunocytochemistry and determination of transepithelial electrical resistance (TEER) before and after sheet detachment and transfer. HCEC layers showed regular morphology with appropriate TEER. Cells were positive for function-associated marker proteins ZO-1, Na+/K+-ATPase, and paxillin, and extracellular matrix proteins fibronectin, laminin and collagen type IV before and after transfer. Sheet detachment and transfer did not impair cell viability. Subsequently, a potential application in ophthalmology was demonstrated by transplantation onto de-endothelialized porcine corneas in vitro. The novel thermo-responsive cell culture carrier facilitates the generation and transfer of functional HCEC sheets. This paves the way to generate tissue engineered human corneal endothelium as an alternative transplant source for endothelial keratoplasty.

Details

Original languageEnglish
Article number045003
Number of pages13
JournalScience and technology of advanced materials
Volume16
Issue number4
Publication statusPublished - Aug 2015
Peer-reviewedYes

External IDs

WOS 000360956800007
Scopus 84940416352
ORCID /0000-0003-3205-1876/work/142256451
ORCID /0000-0003-0189-3448/work/159607167

Keywords

Keywords

  • thermo-responsive polymer, physico-chemical properties, biomolecular functionalization, poly(N-isopropylacrylamide), corneal endothelial cell sheet, tissue engineering, endothelial transplantation, CORNEAL ENDOTHELIAL-CELLS, EXTRACELLULAR-MATRIX, POLYMER SURFACES, CHELATABLE IRON, INTEGRINS, PEPTIDES, SHEETS, RGD, IMMOBILIZATION, METHYL-ETHER)