Human mesenchymal stem cells retain multilineage differentiation capacity including neural marker expression after extended in vitro expansion
Okolicsanyi R.K. Camilleri E.T. Oikari L.E. Yu C. Cool S.M. Van Wijnen A.J. Griffiths L.R. Haupt L.M.
Okolicsanyi R.K.
Camilleri E.T.
Oikari L.E.
Yu C.
Van Wijnen A.J.
Griffiths L.R.
Haupt L.M.
Citations
Altmetric:
Alternative Title
Abstract
The suitability of human mesenchymal stem cells (hMSCs) in regenerative medicine relies on retention of their proliferative expansion potential in conjunction with the ability to differentiate toward multiple lineages. Successful utilisation of these cells in clinical applications linked to tissue regeneration requires consideration of biomarker expression, time in culture and donor age, as well as their ability to differentiate towards mesenchymal (bone, cartilage, fat) or non-mesenchymal (e.g., neural) lineages. To identify potential therapeutic suitability we examined hMSCs after extended expansion including morphological changes, potency (stemness) and multilineage potential. Commercially available hMSC populations were expanded in vitro for > 20 passages, equating to > 60 days and > 50 population doublings. Distinct growth phases (A-C) were observed during serial passaging and cells were characterised for stemness and lineage markers at representative stages (Phase A: P+5, approximately 13 days in culture; Phase B: P+7, approximately 20 days in culture; and Phase C: P+13, approximately 43 days in culture). Cell surface markers, stem cell markers and lineage-specific markers were characterised by FACS, ICC and Q-PCR revealing MSCs maintained their multilineage potential, including neural lineages throughout expansion. Co-expression of multiple lineage markers along with continued CD45 expression in MSCs did not affect completion of osteogenic and adipogenic specification or the formation of neuro-spheres. Improved standardised isolation and characterisation of MSCs may facilitate the identification of biomarkers to improve therapeutic efficacy to ensure increased reproducibility and routine production of MSCs for therapeutic applications including neural repair. Copyright: © 2015 Okolicsanyi et al.
Keywords
5' nucleotidase, beta1 integrin, CD146 antigen, CD200 antigen, CD24 antigen, CD45 antigen, CD56 antigen, endoglin, Hermes antigen, kruppel like factor 4, Thy 1 antigen, transcription factor HES 1, transcription factor PAX3, transcription factor PAX9, transcription factor Sox5, transcription factor Sox6, transcription factor Sox9, biological marker, CD45 antigen, transcription factor, adipocyte, adipogenesis, adult, Article, bone development, cell differentiation, cell expansion, cell growth, cell lineage, controlled study, female, fluorescence activated cell sorting, gene expression, human, human cell, immunocytochemistry, in vitro study, male, mesenchymal stem cell, neural stem cell, osteoblast, polymerase chain reaction, protein expression, quantitative analysis, stem cell culture, young adult, cell culture, cell lineage, cell membrane, cell proliferation, cell shape, cytology, flow cytometry, gene expression profiling, gene expression regulation, mesenchymal stroma cell, metabolism, multicellular spheroid, nerve cell, Western blotting, Adipogenesis, Antigens, CD45, Biomarkers, Blotting, Western, Cell Differentiation, Cell Lineage, Cell Membrane, Cell Proliferation, Cell Shape, Cells, Cultured, Flow Cytometry, Gene Expression Profiling, Gene Expression Regulation, Humans, Mesenchymal Stromal Cells, Neurons, Osteogenesis, Spheroids, Cellular, Transcription Factors
Source Title
PLoS ONE
Publisher
Series/Report No.
Collections
Rights
Attribution 4.0 International
Date
2015
DOI
10.1371/journal.pone.0137255
Type
Article