TY - JOUR
T1 - Epigenetic changes in umbilical cord mesenchymal stromal cells upon stimulation and culture expansion
AU - De Witte, Samantha F.H.
AU - Peters, Fleur S.
AU - Merino, Ana
AU - Korevaar, Sander S.
AU - Van Meurs, Joyce B.J.
AU - O'Flynn, Lisa
AU - Elliman, Steve J.
AU - Newsome, Philip N.
AU - Boer, Karin
AU - Baan, Carla C.
AU - Hoogduijn, Martin J.
N1 - Funding Information: This project has received funding from the European Union's Seventh Framework Programme for Research, technological development and demonstration under grant agreement number 602363 (MEsenchymal stem cells to Reduce Liver INflammation project). Publisher Copyright: © 2018 International Society for Cellular Therapy
PY - 2018/7
Y1 - 2018/7
N2 - Background: Mesenchymal stromal cells (MSCs) are studied for their immunotherapeutic potential. Prior to therapeutic use, MSCs are culture expanded to obtain the required cell numbers and, to improve their efficacy, MSCs may be primed in vitro. Culture expansion and priming induce phenotypical and functional changes in MSCs and thus standardisation and quality control measurements come in need. We investigated the impact of priming and culturing on MSC DNA methylation and examined the use of epigenetic profiling as a quality control tool. Methods: Human umbilical cord–derived MSCs (ucMSCs) were cultured for 3 days with interferon (IFN)γ transforming growth factor (TGF)β or a multi-factor combination (MC; IFNγ TGFβ and retinoic acid). In addition, ucMSCs were culture expanded for 14 days. Phenotypical changes and T-cell proliferation inhibition capacity were examined. Genome-wide DNA methylation was measured with Infinium MethylationEPIC Beadchip. Results: Upon priming, ucMSCs exhibited a different immunophenotype and ucMSC(IFNγ) and ucMSC(MC) had an increased capacity to inhibit T-cell proliferation. DNA methylation patterns were minimally affected by priming, with only one significantly differentially methylated site (DMS) in IFNγ- and MC-primed ucMSCs associated with autophagy activity. In contrast, 14 days after culture expansion, ucMSCs displayed minor phenotypical and functional changes but showed >4000 significantly DMSs, mostly concerning genes involved in membrane composition, cell adhesion and transmembrane signalling. Discussion: These data show that DNA methylation of MSCs is only marginally affected by priming, whereas culture expansion and subsequent increased cellular interactions have a large impact on methylation. On account of this study, we suggest that DNA methylation analysis is a useful quality control tool for culture expanded therapeutic MSCs.
AB - Background: Mesenchymal stromal cells (MSCs) are studied for their immunotherapeutic potential. Prior to therapeutic use, MSCs are culture expanded to obtain the required cell numbers and, to improve their efficacy, MSCs may be primed in vitro. Culture expansion and priming induce phenotypical and functional changes in MSCs and thus standardisation and quality control measurements come in need. We investigated the impact of priming and culturing on MSC DNA methylation and examined the use of epigenetic profiling as a quality control tool. Methods: Human umbilical cord–derived MSCs (ucMSCs) were cultured for 3 days with interferon (IFN)γ transforming growth factor (TGF)β or a multi-factor combination (MC; IFNγ TGFβ and retinoic acid). In addition, ucMSCs were culture expanded for 14 days. Phenotypical changes and T-cell proliferation inhibition capacity were examined. Genome-wide DNA methylation was measured with Infinium MethylationEPIC Beadchip. Results: Upon priming, ucMSCs exhibited a different immunophenotype and ucMSC(IFNγ) and ucMSC(MC) had an increased capacity to inhibit T-cell proliferation. DNA methylation patterns were minimally affected by priming, with only one significantly differentially methylated site (DMS) in IFNγ- and MC-primed ucMSCs associated with autophagy activity. In contrast, 14 days after culture expansion, ucMSCs displayed minor phenotypical and functional changes but showed >4000 significantly DMSs, mostly concerning genes involved in membrane composition, cell adhesion and transmembrane signalling. Discussion: These data show that DNA methylation of MSCs is only marginally affected by priming, whereas culture expansion and subsequent increased cellular interactions have a large impact on methylation. On account of this study, we suggest that DNA methylation analysis is a useful quality control tool for culture expanded therapeutic MSCs.
KW - DNA methylation
KW - culture expansion
KW - epigenetics
KW - mesenchymal stromal cell
KW - priming
KW - quality control
UR - http://www.scopus.com/inward/record.url?scp=85048791644&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.jcyt.2018.05.005
DO - https://doi.org/10.1016/j.jcyt.2018.05.005
M3 - Article
C2 - 29934259
SN - 1465-3249
VL - 20
SP - 919
EP - 929
JO - Cytotherapy
JF - Cytotherapy
IS - 7
ER -