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An in vitro expansion score for tissue-engineering applications with human bone marrow-derived mesenchymal stem cells.
J Tissue Eng Regen Med 2016; 10(2):149-61JT

Abstract

Human bone marrow-derived mesenchymal stem cells (MSCs) have limited growth potential in vitro and cease to divide due to replicative senescence, which from a tissue-engineering perspective has practical implications, such as defining the correct starting points for differentiation and transplantation. Time spent in culture before the loss of required differentiation potential is different and reflects patient variability, which is a problem for cell expansion. This study aimed to develop a score set which can be used to quantify the senescent state of MSCs and predict whether cells preserve their ability to differentiate to osteogenic, adipogenic and chondrogenic phenotypes, based on colony-forming unit (CFU) assay, population doubling time (PDT), senescence-associated β-galactosidase (SA-β-Gal) activity, cell size, telomere length and gene expression of MSCs cultured in vitro over 11 passages. This set of morphological, physiological and genetic senescence markers was correlated to the ability of MSCs to differentiate. Differentiation efficiency was assessed by marker genes and protein expression. CFUs decreased with increasing passage number, whereas SA-β-Gal activity and PDT increased; however, the correlation with MSCs' differentiation potential was sometimes unexpected. The expression of genes related to senescence was higher in late-passage cells than in early-passage cells. Early-passage cells underwent efficient osteogenic differentiation, with mid-passage cells performing best in chondrogenic differentiation. Late-passage cells preserve only adipogenic differentiation potential. Based on this marker set, we propose a senescence score in which combined markers give a reliable quality control of MSCs, not depending only on mechanistic passage number.

Authors+Show Affiliations

Swiss Paraplegic Research, Nottwil, Switzerland. Institute for Surgical Technology and Biomechanics, University of Bern, Switzerland.Swiss Paraplegic Research, Nottwil, Switzerland.Swiss Paraplegic Research, Nottwil, Switzerland.Swiss Paraplegic Research, Nottwil, Switzerland.Swiss Paraplegic Centre, Nottwil, Switzerland. School of Medicine, Griffith University, Brisbane, Queensland, Australia.Cantonal Hospital of Lucerne, Lucerne, Switzerland.Institute for Biomechanics, ETH Zurich, Switzerland.Swiss Paraplegic Research, Nottwil, Switzerland. Institute for Surgical Technology and Biomechanics, University of Bern, Switzerland.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23576360

Citation

Bertolo, Alessandro, et al. "An in Vitro Expansion Score for Tissue-engineering Applications With Human Bone Marrow-derived Mesenchymal Stem Cells." Journal of Tissue Engineering and Regenerative Medicine, vol. 10, no. 2, 2016, pp. 149-61.
Bertolo A, Mehr M, Janner-Jametti T, et al. An in vitro expansion score for tissue-engineering applications with human bone marrow-derived mesenchymal stem cells. J Tissue Eng Regen Med. 2016;10(2):149-61.
Bertolo, A., Mehr, M., Janner-Jametti, T., Graumann, U., Aebli, N., Baur, M., ... Stoyanov, J. V. (2016). An in vitro expansion score for tissue-engineering applications with human bone marrow-derived mesenchymal stem cells. Journal of Tissue Engineering and Regenerative Medicine, 10(2), pp. 149-61. doi:10.1002/term.1734.
Bertolo A, et al. An in Vitro Expansion Score for Tissue-engineering Applications With Human Bone Marrow-derived Mesenchymal Stem Cells. J Tissue Eng Regen Med. 2016;10(2):149-61. PubMed PMID: 23576360.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - An in vitro expansion score for tissue-engineering applications with human bone marrow-derived mesenchymal stem cells. AU - Bertolo,Alessandro, AU - Mehr,Marco, AU - Janner-Jametti,Tiziana, AU - Graumann,Ursula, AU - Aebli,Niklaus, AU - Baur,Martin, AU - Ferguson,Stephen J, AU - Stoyanov,Jivko V, Y1 - 2013/04/10/ PY - 2012/07/09/received PY - 2013/01/08/revised PY - 2013/01/30/accepted PY - 2013/4/12/entrez PY - 2013/4/12/pubmed PY - 2016/11/1/medline KW - adipogenesis KW - chondrogenesis KW - in vitro senescence KW - mesenchymal stem cells KW - osteogenesis SP - 149 EP - 61 JF - Journal of tissue engineering and regenerative medicine JO - J Tissue Eng Regen Med VL - 10 IS - 2 N2 - Human bone marrow-derived mesenchymal stem cells (MSCs) have limited growth potential in vitro and cease to divide due to replicative senescence, which from a tissue-engineering perspective has practical implications, such as defining the correct starting points for differentiation and transplantation. Time spent in culture before the loss of required differentiation potential is different and reflects patient variability, which is a problem for cell expansion. This study aimed to develop a score set which can be used to quantify the senescent state of MSCs and predict whether cells preserve their ability to differentiate to osteogenic, adipogenic and chondrogenic phenotypes, based on colony-forming unit (CFU) assay, population doubling time (PDT), senescence-associated β-galactosidase (SA-β-Gal) activity, cell size, telomere length and gene expression of MSCs cultured in vitro over 11 passages. This set of morphological, physiological and genetic senescence markers was correlated to the ability of MSCs to differentiate. Differentiation efficiency was assessed by marker genes and protein expression. CFUs decreased with increasing passage number, whereas SA-β-Gal activity and PDT increased; however, the correlation with MSCs' differentiation potential was sometimes unexpected. The expression of genes related to senescence was higher in late-passage cells than in early-passage cells. Early-passage cells underwent efficient osteogenic differentiation, with mid-passage cells performing best in chondrogenic differentiation. Late-passage cells preserve only adipogenic differentiation potential. Based on this marker set, we propose a senescence score in which combined markers give a reliable quality control of MSCs, not depending only on mechanistic passage number. SN - 1932-7005 UR - https://www.unboundmedicine.com/medline/citation/23576360/An_in_vitro_expansion_score_for_tissue_engineering_applications_with_human_bone_marrow_derived_mesenchymal_stem_cells_ L2 - https://doi.org/10.1002/term.1734 DB - PRIME DP - Unbound Medicine ER -