Improved Post-Thaw Function and Epigenetic Changes in Mesenchymal Stromal Cells Cryopreserved Using Multicomponent Osmolyte Solutions

Kathryn Pollock, Rebekah M. Samsonraj, Amel Dudakovic, Roman Thaler, Aron Stumbras, David H. McKenna, Peter I. Dosa, Andre J van Wijnen, Allison Hubel

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Current methods for freezing mesenchymal stromal cells (MSCs) result in poor post-Thaw function, which limits the clinical utility of these cells. This investigation develops a novel approach to preserve MSCs using combinations of sugars, sugar alcohols, and small-molecule additives. MSCs frozen using these solutions exhibit improved post-Thaw attachment and a more normal alignment of the actin cytoskeleton compared to cells exposed to dimethylsulfoxide (DMSO). Osteogenic and chondrogenic differentiation assays show that cells retain their mesenchymal lineage properties. Genomic analysis indicates that the different freezing media evaluated have different effects on the levels of DNA hydroxymethylation, which are a principal epigenetic mark and a key step in the demethylation of CpG doublets. RNA sequencing and quantitative real time-polymerase chain reaction validation demonstrate that transcripts for distinct classes of cytoprotective genes, as well as genes related to extracellular matrix structure and growth factor/receptor signaling are upregulated in experimental freezing solutions compared to DMSO. For example, the osmotic regulator galanin, the antiapoptotic marker B cell lymphoma 2, as well as the cell surface adhesion molecules CD106 (vascular cell adhesion molecule 1) and CD54 (intracellular adhesion molecule 1) are all elevated in DMSO-free solutions. These studies validate the concept that DMSO-free solutions improve post-Thaw biological functions and are viable alternatives for freezing MSCs. These novel solutions promote expression of cytoprotective genes, modulate the CpG epigenome, and retain the differentiation ability of MSCs, suggesting that osmolyte-based freezing solutions may provide a new paradigm for therapeutic cell preservation.

Original languageEnglish (US)
Pages (from-to)828-842
Number of pages15
JournalStem Cells and Development
Volume26
Issue number11
DOIs
StatePublished - Jun 1 2017

Fingerprint

Mesenchymal Stromal Cells
Epigenomics
Freezing
Dimethyl Sulfoxide
Sugar Alcohols
RNA Sequence Analysis
Galanin
Growth Factor Receptors
Vascular Cell Adhesion Molecule-1
Cell Adhesion Molecules
B-Cell Lymphoma
Actin Cytoskeleton
Genes
Extracellular Matrix
Real-Time Polymerase Chain Reaction
Gene Expression
DNA
Therapeutics

Keywords

  • amino acids

ASJC Scopus subject areas

  • Hematology
  • Developmental Biology
  • Cell Biology

Cite this

Pollock, K., Samsonraj, R. M., Dudakovic, A., Thaler, R., Stumbras, A., McKenna, D. H., ... Hubel, A. (2017). Improved Post-Thaw Function and Epigenetic Changes in Mesenchymal Stromal Cells Cryopreserved Using Multicomponent Osmolyte Solutions. Stem Cells and Development, 26(11), 828-842. https://doi.org/10.1089/scd.2016.0347

Improved Post-Thaw Function and Epigenetic Changes in Mesenchymal Stromal Cells Cryopreserved Using Multicomponent Osmolyte Solutions. / Pollock, Kathryn; Samsonraj, Rebekah M.; Dudakovic, Amel; Thaler, Roman; Stumbras, Aron; McKenna, David H.; Dosa, Peter I.; van Wijnen, Andre J; Hubel, Allison.

In: Stem Cells and Development, Vol. 26, No. 11, 01.06.2017, p. 828-842.

Research output: Contribution to journalArticle

Pollock, K, Samsonraj, RM, Dudakovic, A, Thaler, R, Stumbras, A, McKenna, DH, Dosa, PI, van Wijnen, AJ & Hubel, A 2017, 'Improved Post-Thaw Function and Epigenetic Changes in Mesenchymal Stromal Cells Cryopreserved Using Multicomponent Osmolyte Solutions', Stem Cells and Development, vol. 26, no. 11, pp. 828-842. https://doi.org/10.1089/scd.2016.0347
Pollock, Kathryn ; Samsonraj, Rebekah M. ; Dudakovic, Amel ; Thaler, Roman ; Stumbras, Aron ; McKenna, David H. ; Dosa, Peter I. ; van Wijnen, Andre J ; Hubel, Allison. / Improved Post-Thaw Function and Epigenetic Changes in Mesenchymal Stromal Cells Cryopreserved Using Multicomponent Osmolyte Solutions. In: Stem Cells and Development. 2017 ; Vol. 26, No. 11. pp. 828-842.
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