Journal Article

Human Mesenchymal Stem Cells Retain Multilineage Differentiation Capacity Including Neural Marker Expression after Extended In Vitro Expansion

Cool, Simon M., Okolicsanyi, Rachel K., Oikari, Lotta E, Yu, Chieh, Griffiths, Lyn R., van Wijnen, Andre J., Camilleri, Emily T., Haupt, Larisa M.

2015

Overview

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 neurospheres. 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.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Adipogenesis / Biochemistry / Biocompatibility / Biomarkers / Biomarkers - metabolism / Biomedical materials / Blotting, Western / Bone marrow / Bone surgery / Cartilage / CD45 antigen / Cell culture / Cell Differentiation / Cell Lineage / Cell Membrane - metabolism / Cell Proliferation / Cell Shape / Cell surface / Cells, Cultured / Expansion / Flow Cytometry / Gene Expression Profiling / Gene Expression Regulation / Genetic aspects / Genetic markers / Genomics / Health aspects / Homeostasis / Humans / Leukocyte Common Antigens - metabolism / Mesenchymal stem cells / Mesenchymal Stem Cells - cytology / Mesenchyme / Metastasis / Molecular biology / Neurons - cytology / Neurospheres / Orthopedics / Osteogenesis / Physiological aspects / Regeneration (physiology) / Regenerative medicine / Reproducibility / Spheroids, Cellular - cytology / Stem cells / Surface markers / Therapeutic applications / Tissue engineering / Transcription Factors - metabolism / Tumors