Cell encapsulation and cryostorage in PVA-gelatin cryogels: Incorporation of carboxylated ε-poly-L-lysine as cryoprotectant

Nihal E. Vrana, Kazuaki Matsumura, Suong Hyu Hyon, Luke M. Geever, James E. Kennedy, John G. Lyons, Clement L. Higginbotham, Paul A. Cahill, Garrett B. McGuinness

Research output: Contribution to journalArticlepeer-review

30 Citations (Scopus)

Abstract

It is desirable to produce cryopreservable cell-laden tissue-engineering scaffolds whose final properties can be adjusted during the thawing process immediately prior to use. Polyvinyl alcohol (PVA)-based solutions provide platforms in which cryoprotected cell suspensions can be turned into a ready-to-use, cell-laden scaffold by a process of cryogelation. In this study, such a PVA system, with DMSO as the cryoprotectant, was successfully developed. Vascular smooth muscle cell (vSMC)-encapsulated cryogels were investigated under conditions of cyclic strain and in co-culture with vascular endothelial cells to mimic the environment these cells experience in vivo in a vascular tissue-engineering setting. In view of the cytotoxicity DMSO imposes with respect to the production procedure, carboxylated poly-L-lysine (COOH-PLL) was substituted as a non-cytotoxic cryoprotectant to allow longer, slower thawing periods to generate more stable cryogels. Encapsulated vSMC with DMSO as a cryoprotectant responded to 10% cyclic strain with increased alignment and proliferation. Cells were stored frozen for 1 month without loss of viability compared to immediate thawing. SMC-encapsulated cryogels also successfully supported functional endothelial cell co-culture. Substitution of COOH-PLL in place of DMSO resulted in a significant increase in cell viability in encapsulated cryogels for a range of thawing periods. We conclude that incorporation of COOH-PLL during cryogelation preserved cell functionality while retaining fundamental cryogel physical properties, thereby making it a promising platform for tissue-engineering scaffolds, particularly for vascular tissue engineering, or cell preservation within microgels.

Original languageEnglish
Pages (from-to)280-290
Number of pages11
JournalJournal of Tissue Engineering and Regenerative Medicine
Volume6
Issue number4
DOIs
Publication statusPublished - Apr 2012

Keywords

  • Cell encapsulation
  • Cryogel
  • Cryoprotectant
  • Cryostorage
  • Cyclic strain
  • Gelatin
  • PLL
  • PVA

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